1.Effects of RNA Interfering of MBP-1 on Proliferation of Saos-2 Cell Line
Xinhe SHI ; Zhe GENG ; Xingchen SHI ; Kejun MA ; Hongwen ZHU ; Wen REN ; Yali ZHOU
Journal of China Medical University 2016;45(7):604-609
Objective To investigate the effects of c?myc promoter binding protein 1(MBP?1)gene on the proliferation of human Saos?2 osteo?sarcoma cells in vitro. Methods Saos?2 cells were divided into three groups:blank control group(untransfected cells),negative group(cells transfected with missense sequence)and experimental group(cells transfected with MBP?1 shRNA). Two MBP?1 shRNA sequences and one neg?ative control shRNA sequence were designed ,synthesized and cloned into pSIREN?retroQ plasma. Then the recombinant plasmids were construct?ed and transfected into human Saos?2 osteosarcoma cells by Lipofectamine 2000. The expressions of MBP?1 mRNA and protein in Saos?2 cells were detected by real?time PCR and Western blot ,respectively. The effects of altered expression of MBP?1 on cell proliferation were measured by CCK?8 cell proliferation assay. The expressions of cyclin D1 and cyclin E in Saos?2 were determined by Western blot. Results PCR and sequenc?ing results indicated that the recombinant plasmids pSIREN?retroQ was constructed. The relative expression level of MBP?1 mRNA in the MBP?1 siRNA transfection group was significantly decreased than that in blank control group(P<0.05). Compared with the blank control group,the ex?pression levels of MBP?1 protein in the experimental group also significantly decreased. The proliferation abilities of Saos?2 cells at 48,72,and 96 hours after MBP?1 siRNA transfection were significantly increased than those in the blank control group(P<0.05). Compared with the blank con?trol group,the expression levels of cyclin D1 and cyclin E protein in the experimental group also significantly increased(P<0.05). Conclusion Knockdown of the expression of MBP?1 gene promotes the proliferation of human Saos?2 osteosarcoma cells. MBP?1 gene may become the new tar?get of gene therapy for osteosarcoma.
2.Construction of shSet7/9 vector and its function in HepG2
Kejun MA ; Xingchen SHI ; Ping LI ; Xiaoqiang LI ; Wen REN ; Long QIN ; Xinhe SHI
Journal of Xi'an Jiaotong University(Medical Sciences) 2015;(6):753-757
Objective To silence human gene Set7/9 and screen out stable transfection cell line in hepatocellular carcinoma cell line HepG2 so as to investigate the impact of down-regulation of Set7/9 in cell line HepG2 and provide experimental foundation for studies on the effect of set7/9 in HepG2.Methods The target oligo was designed and synthesized;shRNA interference vector and the control vector were constructed and transfected into HepG2 cells;the stable transfection cells were screened out.Then Real-time PCR and Western blot were performed to detect the silence of Set7/9 according to both gene expression and protein expression level. Results The shRNA interference vector was constructed and transfected into HepG2 cells successfully.Compared with that in the negative control group,the expression of Set7/9 was dramatically downregulated (P < 0.05 ). Meanwhile,the expression of related protein Sirt1 and Suv39h1 was upregulated 8.4 folds and 1.1 fold, respectively.Conclusion Downregulation of Set7/9 expression can upregulate Sirt1 and Suv39h1,suggesting that Set7/9 may affect the activity of HepG2 cell lines.
3.Preparation and performance evaluation of a novel detrgent-enzymatic tissue-engineered tracheal matrix in rabbits
Fei SUN ; Shu PAN ; Hongcan SHI ; Fangbiao ZHANG ; Weidong ZHANG ; Gang YE ; Xingchen LIU
Chinese Journal of Thoracic and Cardiovascular Surgery 2014;30(1):38-41,55
Objective To determine the detergent-enzymatic cycles and evaluate the biomechanical characteristics as well as extracellular matrix integrity of the decellularized tracheal scaffold in rabbit.Methods Forty tracheal segments were harvested from New Zealand white rabbits.Thirty-five of tracheas were subjected to a detergent-enzymatic method of decellularization for 1/3/5/6/7/8/9 cycles,respectively,and other five were stored in phosphate-buffered saline at 4℃ as a control.Comparative examinations were performed by the macroscopic view,histological view(hematoxylin and eosin stain,Movat Pentachrome stain,4-6-diamidino-2-phenylindole),scanning electron microscope (SEM) and biomechanical properties between decellularized groups and control group.Results After 7 detergent-enzymatic cycles,almost complete decellularized tracheae,retaining the hierarchical and mechanical properties of the native tissues,could be obtained.Histological and molecular biology analysis demonstrated that all cellular components and nuclear material were removed.SEM analysis revealed that the decellularized matrices retained the hierarchical structures of native trachea,and biomechanical tests showed that decellularization approach did not led to any influence on tracheal morphological and mechanical properties.Immunofluorescence analysis show a significant reduction of nuclear material in decellularized tracheas (P < 0.05).Conclusion In conclusion,this work suggests that 7 cycles of the modified DEM generates a bioengineered rabbit tracheal matrix that is structurally and mechanically similar to native trachea which could be a better selection for tracheal reconstruction with tissue engineering method.
4.Advanced glycosylation end products promote migration of podocytes through mTORC1/uPAR pathway
Xiaofan TAN ; Yuanhan CHEN ; Chunping YU ; Yuxiong LAI ; Li ZHANG ; Xingchen ZHAO ; Hong ZHANG ; Ting LIN ; Ruizhao LI ; Wei SHI
Chinese Journal of Pathophysiology 2014;(12):2232-2237
[ ABSTRACT] AIM: To investigate the influence of advanced glycosylation end products-modified bovine serum albumin (AGE-BSA) on mammalian target of rapamycin complex 1 (mTORC1), urokinase-type plasminogen activator re-ceptor ( uPAR) , and cell mobility in the podocytes, and to further explore the probable relationship.METHODS: The conditionally immortalized mouse podocyte cell line was cultured in vitro.MTT assay and immunofluorescence were used to analyze the cell viability and cytoskeleton of the podocytes treated with the stimuli and intervention agents.The activity of mTORC1 and the expression level of uPAR in normal podocytes and podocytes treated with control BSA or AGE-BSA were detected by Western blotting.The migration ability of the podocytes was determined by would-healing assay.Rapamycin was added to inhibit the activity of mTORC1 along with the addition of AGE-BSA to observe the changes of uPAR and the motility of podocytes.RESULTS:No significant difference of the cell viability or cytoskeleton in the podocytes treated with the stimuli and intervention agents was observed.AGE-BSA up-regulated the activity of mTORC1 and the expression of uPAR, and induced the high mobility of the podocytes.Rapamycin obviously reduced the high expression level of uPAR and the increase in the migration ability of podocytes caused by AGE-BSA treatment.CONCLUSION: AGE-BSA might cause the high migration of podocytes through the mTORC1/uPAR signaling pathway.
5.Diagnosis of small round cell tumor of nasal sinus and bases of skull (a report of 123 cases).
Wenbin LEI ; Zhenzhong SU ; Weiping WEN ; Jianbo SHI ; Bing LIAO ; Xingchen WU ; Geng XU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2008;22(20):929-933
OBJECTIVE:
To investigate the features and diagnosis way of small round cell tumor of nasal sinus and bases of skull.
METHOD:
A retrospective analysis of 123 case with small round cell tumor of nasal sinus and bases of skull were carried out in our hospital in past ten year. Clinical, histological, radiological and immunohistochemical characters of these cases were studied.
RESULT:
All cases usually complained of nasal obstruction, headache, diplopia, nasal mucus with bleeding, vision or weight loss. Expansible or infiltrative lumps were found in nasal sinus or bases of skull in radiological examination. A lot of small round cells were found in these tumors in histological pathology. At least 5-6 cell, tissue or tumor markers were examined immunohistochemically in most of cases before the final diagnosis were made. In some cases over 20 markers were examined. Five cases were carried out transmission electron microscope examination, special features such as desmosome and myofilament were found.
CONCLUSION
Clinical symptom, physical signs and radiological finding can supply malignant evidences of these tumors. Histological examination can make certain that they are small round cell tumors, but final diagnosis is still hard to make only by these. Immunohistochemical examination of various markers can tell the original characters of the specimen tissues, it is the key for final diagnosis. Transmission electron microscope examination is another helpful way for diagnosis.
Adolescent
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Adult
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Aged
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Aged, 80 and over
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Child
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Child, Preschool
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Humans
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Infant
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Leukemia, Lymphocytic, Chronic, B-Cell
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diagnosis
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pathology
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Male
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Middle Aged
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Paranasal Sinus Neoplasms
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diagnosis
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pathology
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Retrospective Studies
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Skull Base Neoplasms
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diagnosis
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pathology
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Young Adult
6.Analysis of risk factors for multiple myeloma combined with radiation-based sarcopenia
Jie REN ; Lei QI ; Xingchen YAO ; Jinzhou WANG ; Xiangjun SHI ; Wenming CHEN ; Xinru DU
Chinese Journal of Orthopaedics 2023;43(9):567-573
Objective:To analyze the risk factors of radiation-based sarcopenia in patients with multiple myeloma (MM).Methods:A total of 185 clinical and imaging data of patients with MM admitted to Beijing Chaoyang Hospital from September 2009 to October 2019 were retrospectively analyzed. The area of the erector spinae muscle and the area of fatty infiltration (FI) in the fascial compartment were measured by Image-pro Ρlus software, and the area of the fat-free erector spinae muscle and the fat infiltration rate (FI%) were calculated. Sarcopenia was defined as an erector spinae area of less than 3 197 mm 2 in males and 2 895 mm 2 in females. The differences in gender, age, body mass index, disease duration, hemoglobin, leukocytes, platelets, albumin, serum calcium, lactate dehydrogenase, serum creatinine, alkaline phosphatase, M-protein, serum β 2-microglobulin, bortezomib chemotherapy, receipt of stem cell transplantation, osteopathy, stage, recurrence and progression of MM between the sarcopenia group and the normal muscle group were compared. Binary logistic regression was used to analyze the independent risk factors of sarcopenia in MM patients. Kaplan-Meier curves were drawn to compare the survival rates between the two groups. Results:53.0% (98/185) of MM patients were complicated with sarcopenia: there were 30 males, whose fat-free erector spinae area was 25.0±6.0 cm 2, the FI of erector spinae was 12.0±4.8 cm 2, and the FI% was 31.5%±12.0%, while there were 68 females, whose fat-free erector spinae area was 22.7±4.2 cm 2, the FI of erector spinae was 10.7±4.1 cm 2, and the FI% was 30.2%±9.8%. 47.0% (87/185) of MM patients had normal muscle mass: there were 62 males, whose fat-free erector spinae area was 40.6±6.5 cm 2, the FI of erector spinae was 9.3±4.8 cm 2, and the FI% was 17.9%±7.4%, while there were 25 females, whose fat-free erector spinae area was 33.6±5.1 cm 2, the FI of erector spinae was 9.9±3.0 cm 2, and the FI% was 21.9%±5.7%. There were statistically significant differences in the gender composition ratio (χ 2=30.47, P<0.001), hemoglobin ( t=-2.73, P=0.007), serum creatinine ( Z=-2.26, P=0.024), receipt of stem cell transplantation (χ 2=4.32, P=0.038), and MM recurrence and progression (χ 2=3.85, P=0.050) between the two groups. However, there were no significant differences in age, body mass index, course of disease, leukocytes, platelets, albumin, alkaline phosphatase, lactate dehydrogenase, serum calcium, M-protein, serum β 2-microglobulin, bortezomib chemotherapy, osteopathy or MM stage ( P>0.05). Binary logistic regression analysis showed that female was an independent risk factor for sarcopenia in MM patients. The survival rates at 2, 3, 4, and 5 years were 87.9%, 71.8%, 64.4%, and 53.7% in the sarcopenia group, and 92.1%, 75.8%, 66.8%, and 66.8% in the normal muscle group, respectively, with no statistically significant differences ( HR=0.71, P=0.364). Conclusion:The incidence of radiation-based sarcopenia in MM patients is 53.0%. Low hemoglobin and blood creatinine levels, not receiving stem cell transplantation, and recurrence or progression of MM are associated with sarcopenia in MM patients, and female is an independent risk factor for sarcopenia in MM patients.
7.Prefrontal cortex activation in women with perimenopausal depression: a functional near-infrared spectroscopy study
Cancan YU ; Jiao SHI ; Zhengxian XU ; Xiaodong SONG ; Ziyi WU ; Xingchen ZHOU ; Mengyang WANG ; Dan WANG ; Shangjie CHEN ; Min WANG
Chinese Journal of Behavioral Medicine and Brain Science 2023;32(10):894-900
Objective:To explore the relative concentration changes of oxygenated hemoglobin (Oxy Hb) in the prefrontal cortex (PFC) and brain region activation during emotional face recognition tasks in women with perimenopausal depression.Methods:From February to April 2023, forty perimenopausal women were recruited, including 20 women with perimenopausal depression (experimental group) and 20 women with non-perimenopausal depression (control group). All participants were evaluated by the modified Kupperman score, 24-item Hamilton depression scale (HAMD-24), and patient health questionnaire (PHQ-9). Functional near-infrared spectroscopy (fNIRS) equipment was used to measure the relative concentration of Oxy-Hb in the PFC in two groups under the emotional face recognition task. Statistical analysis was performed by SPSS 26.0 software. Data were analyzed by a t-test, rank sum test, and Pearson correlation. Results:There were statistically significant differences in the results of the modified Kupperman score((23.20±3.66), (18.10±1.28)), HAMD-24((15.95±5.47), (3.35±1.84)), and PHQ-9(7.00(5.00, 10.75), 1.50(1.00, 3.00)) scales between the the experimental group and control group ( P<0.05). There was a positive correlation between the modified Kupperman score and the HAMD-24 score in the experimental group ( r=0.685, P=0.01). The reaction time of the experimental group in identifying negative and neutral emotional faces was statistically significant compared to the control group( t=4.01, 4.80, both P<0.05). Compared with identifying neutral emotions, PFC activation was stronger in the experimental group and control group when identifying negative emotions ( P<0.05). The PFC activation in the experimental group was stronger than that in the control group when identifying negative emotions ( P<0.05). There was no statistically significant difference in the activation level between the two groups when identifying neutral emotions ( P>0.05). Conclusion:Women with perimenopausal depression exhibit specificity in emotional processing, with increased PFC activation when identifying negative emotions, impaired emotional processing function of PFC, and dysfunction of aerobic metabolism.
8.Effects of electro-acupuncture on hippocampal CXCR4 and IP-10 protein expression in rats with depression disorder
Xingchen LIANG ; Huili JIANG ; Danmei ZHANG ; Rongxing SHI ; Yu WANG ; Jing LI ; Xinjing YANG ; Bingcong ZHAO ; Qiuyun YU ; Xiang LI ; Shixing FENG ; Ya TU
Journal of Beijing University of Traditional Chinese Medicine 2017;40(10):872-875
Objective To investigate the effect of electro-acupuncture (EA) on hippocampal chemokine cys-x-cys receptor 4 (CXCR4) and interferon-inducible protein-10 (IP-10) in rats with depression disorder.Methods The rat model of depression disorder was induced by unpredictable chronic-mild stimulation.Forty male rats were randomly divided into normal,model,EA and fluoxetine groups.Rats in EA group received electro-acupuncture at GV20 (Baihui) and GV29 (Yintang).Rats in fluoxetine group were administered with fluoxetine by oral gavage.On the 28th day,the expression of hippocampal CXCR4 and IP-10 were detected by using antibody microarrays technology.Fold change and ratio of the density of the proteins were used to investigate alteration of the differential protein expression.Results Compared with the normal group,unpredictable chronic-mild stimulation up-regulated the expression of hippocampal CXCR4 and IP-10 in model group (fold change =1.21,1.50).Compared with the model group,EA and fluoxetine reversed the up-regulation of hippocampal CXCR4 and IP-10 (EA group:fold change =0.65,0.72;fluoxetine group:fold change =0.57,0.62).Conclusion Effects of electro-acupuncture on depression disorder may be achieved by regulating the expression of hippocampal CXCR4 and IP-10.
9.Survey of the prevalence and epidemiological characteristics of primary palmar hyperhidrosis among adolescents in Yangzhou
YANG Junfeng ; WANG Yao ; SHAN Yibo ; LIU Xingchen ; FAN Yiwei ; SHI Hongcan
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2017;24(10):774-778
Objective To investigate the prevalence and related factors of primary palmar hyperhidrosis in adolescents in Yangzhou. Methods On-site questionnaire survey was performed on students selected by cluster random sampling from the two colleges and two high or middle schools, with each class as a unit. Data were collected through the questionnaire to make the diagnosis and severity grading. Results A total of 3 487 copies of the questionnaire were distributed in the survey and 3 299 were finished, among which 3 083 were effective with an effective rate of 88.41%. Among them, 1 358 respondents were males and 1 725 were females; 933 were middle school students, 809 high school students, and the remaining 1 341 college students. According to the diagnostic criteria, 104 respondents were diagnosed with palmar hyperhidrosis with an overall prevalence of 3.37%. There were 60 (4.41%) males and 44 (2.55%) females. Although the prevalence of palmar hyperhidrosis in males was higher than that of females (χ2=8.130,P<0.05), severe palmar hyperhidrosis was more often to be observed in females than in males, and females were also more likely to have hyperhidrosis in other parts of the body. In addition, the age of the first onset of the disease was mainly 10 to 20 years old and 36.54% of the patients had a family history. Conclusion The prevalence of palmar hyperhidrosis in adolescents in Yangzhou was 3.37%, and there is a significant difference in the gender. The palmar hyperhidros is often accompanied by hyperhidrosis symptoms of other parts of body, and the disease shows an obvious genetic predisposition.