1.Effects of PRMT5 expression on cell proliferation of ovarian cancer cell HO8910
Ye WEI ; Ran-Ran WANG ; Tian-Tian HAN ; Kang-Rong ZHAO ; Qiong LIN ; Gen-Bao SHAO
Journal of Medical Postgraduates 2018;31(6):565-572
Objective Protein arginine methyltransferase 5 (PRMT5),a member of the histone arginine methylation transferase family,is involved in a wide range of biological regulation through ei-ther epigenetic or posttranslational methylation modifications. The pur-pose of the present study was to investigate the effects of PRMT5 on cell proliferation of ovarian cancer cell HO8910. Methods Cell lines HO8910 with PRMT5 overexpression were obtained by transi-ent transfection,which were divided into three groups in the experiment: blank control group (wild-type cell line HO8910),negative control group (HO8910 cells were transfected with pCMV-myc plasmid),and experimental group (HO8910 cells were transfected with pCMV-myc-PRMT5 plasmid). Western blot was used to detect the expression of myc protein,and qRT-PCR was used to detect the ex-pression of PRMT5 mRNA. Cell lines HO8910 with inducible stable knockdown of PRMT5 were established by shRNA interference method,which were divided into four groups: pLKO control group (infected by empty vector lentivirus),pLKO+Dox (100ng/mL) group,shPRMT5 group (infected by PRMT5shRNA lentivirus) and shPRMT5+Dox (100 ng/mL) group. Western blot and qRT-PCR were used to detect the expression of PRMT5 protein and mRNA levels. Dox-induced PRMT5 knockdown was detected by increasing Dox concentration,which includes four groups,Dox 0ng/mL group,Dox 1ng/mL group,Dox 10ng/mL group,Dox 100ng/mL group,and each group was treated for 48 hours. Western blot and qRT-PCR were used to detect the PRMT5 protein and mRNA expression. Colony formation assay,EdU assay,and CCK-8 assay were used to test cells proliferation. The experiment was conducted in two large groups each with two subgroups: PRMT5 knockdown group (Dox-group,Dox+ group),PRMT5 overexpression group (pCMV-myc group,pCMV-myc-PRMT5 group). Western blot was used to detect the effects of PRMT5 expression on proliferation-related proteins. The experiment was conducted in two large groups,PRMT5 knockdown group with four subgroups : Dox 0ng/mL group,Dox 1ng/mL group,Dox 10ng/mL group and Dox 100ng/mL group,and PRMT5 overexpression group with two subgroups (pCMV-myc group and pCMV-myc-PRMT5 group). Results Western blot results showed that the expression of myc was detected in the experimental group in which HO8910 cells were transfected with pCMV-myc-PRMT5,and the expression of PRMT5 mRNA was significantly higher in the experimental group than those in the blank control group and the negative control group (P<0.001) . Western blot and qRT-PCR showed that PRMT5 protein (0.32±0.25) and mRNA expression levels in shPRMT5+Dox group were significantly lower than those of shPRMT5 group (0.89±0.18) (P<0.05). Western blot and qRT-PCR confirmed that PRMT5 protein (0.21±0.24) and mRNA expres-sion in Dox 10ng/mL group and Dox 100ng/mL group (0.08±0.15) were significantly downregulated compared to Dox 0ng/mL group (1.11±0.15) (P<0.05). Colony formation experiments,EdU experiments,and CCK-8 experiments confirmed that the proliferative ca-pacity of cells in Dox+group was lower than that of Dox-group in PRMT5 knockdown group(P<0.05); while in PRMT5 overexpression group,the proliferative capacity of pCMV-myc-PRMT5 group was significantly higher than that of the pCMV-myc group (P<0.05). Western blot results showed that the protein expression of Cyclin D1 was significantly lower in Dox 100 ng/mL group (0.17±0.06) than that in Dox 0 ng/mL group (1.18±0.18) (P<0.05) and the expression of P21 was significantly increased in PRMT5 knockdown group (P<0.05). In the PRMT5 overexpression group,the protein expression of Cyclin D1 in pCMV-myc-PRMT5 group (3.48± 0.22) was higher than that in pCMV-myc group (0.88±0.15) (P<0.05),while the protein expression of P21 (0.08±0.17) were significantly lower than that of pCMV-myc group (4.12±0.10) (P<0.05). Conclusion PRMT5 plays an important role in the proliferation of ovarian cancer cells. Down-regulation of PRMT5 can inhibit cell proliferation and up-regulation of PRMT5 can pro-mote cell proliferation.
3.Establishment of the Mathematical Model for PMI Estimation Using FTIR Spec-troscopy and Data Mining Method
Lei WANG ; Xin-Chao QIN ; Han-Cheng LIN ; Kai-Fei DENG ; Yi-Wen LUO ; Qi-Ran SUN ; Qiu-Xiang DU ; Zhen-Yuan WANG ; Ya TUO ; Jun-Hong SUN
Journal of Forensic Medicine 2018;34(1):1-6,12
Objective To analyse the relationship between Fourier transform infrared (FTIR) spectrum ofrat's spleen tissue and postmortem interval (PMI) for PMI estimation using FTIR spectroscopy combinedwith data mining method. Methods Rats were sacrificed by cervical dislocation, and the cadavers were placed at 20 ℃. The FTIR spectrum data of rats' spleen tissues were taken and measured at different time points. After pretreatment, the data was analysed by data mining method. Results The absorption peak intensity of rat's spleen tissue spectrum changed with the PMI, while the absorption peak position was unchanged. The results of principal component analysis (PCA) showed that the cumulative contribution rate of the first three principal components was 96%. There was an obvious clustering tendency for the spectrum sample at each time point. The methods of partial least squares discriminant analysis (PLS- DA) and support vector machine classification (SVMC) effectively divided the spectrum samples with different PMI into four categories (0-24 h, 48-72 h, 96-120 h and 144-168 h). The determination coefficient (R2) of the PMI estimation model established by PLS regression analysis was 0.96, and the root mean square error of calibration (RMSEC) and root mean square error of cross validation (RMSECV) were 9.90 h and 11.39 h respectively. In prediction set, the R2 was 0.97, and the root mean square error of prediction (RMSEP) was 10.49 h. Conclusion The FTIR spectrum of the rat's spleen tissue can be effectively analyzed qualitatively and quantitatively by the combination of FTIR spectroscopy and data mining method, and the classification and PLS regression models can be established for PMI estimation.
4.A correlation study between intravoxel incoherent motion MRI and histopathology in a hepatocellular carcinoma tumor model
Shuohui YANG ; Fang LU ; Ran GUO ; Zhihong HAN ; Caixia FU ; Menglong ZHAO ; Xueqin BAI ; Jiang LIN
Chinese Journal of Radiology 2018;52(7):549-553
Objective To explore the correlations between intravoxel incoherent motion (IVIM) parameters measured at different time points and histopathological markers in an orthotopic xenograft hepatocellular carcinoma (HCC) nude mice model. Methods A total of 40 HCC orthotopic bearing mouse models were established. When they grew to 21 days, 10 HCC-bearing mice were randomly selected as the baseline group (Group A) by a numeric table method. Then the rest mice were randomly selected on the 28th day, 35th day, and 42nd day of the growth by using the same method, 10 each for B, C, and D groups, respectively. All mice underwent MR IVIM study and apparent diffusion coefficient (ADC), true diffusion coefficient (D), pseudo-diffusion coefficient (D*) and perfusion fraction (f) were measured. After the MRI scanning, the tumors were removed for pathological examination. The necrosis score (NF), tumor size and microvessel density (MVD) were calculated. The IVIM parameters were compared among these 4 groups by Kruskal-Wallis H test and the correlations between these IVIM parameters and histological features were studied with Spearman rank correlation test. Results One tumor in each of C and D groups was excluded because f values of IVIM were close to zero. There were significant differences found in ADC and D among all the 4 groups (P<0.05). However, no difference was found in D*and f (P>0.05). Compared with the baseline (group A), ADC decreased significantly at 7 and 14 days, whilst D decreased significantly at 7 days. The differences in tumor size, MVD and NF between the 4 groups were statistically significant. Compared with the baseline, the tumor size and NF significantly increased at 7, 14 and 21 days, and MVD increased at 14 and 21 days. Significantly positive correlations were demonstrated between ADC and MVD, NF (r=0.461 and 0.442, P<0.05), between D and MVD, NF (r=0.568 and 0.519, P<0.05) after exclusion of the data from the baseline. The parameter f from all the time points including the baseline was positively correlated with histological MVD and NF (r=0.590 and 0.458, P<0.05). Conclusion IVIM parameters may reflect the intratumoral vascularity, tumor cell proliferation and necrosis of HCC, and they are correlated with the pathological indicators.
5.Study on the situation of seat belt wearing among drivers and front-seat passengers of vehicles in Nanjing in 2005 - 2007
Yu QIN ; Ming WU ; Jie YANG ; Jin-Yi ZHOU ; Quan-Yong XIANG ; Ran TAO ; Ren-Qiang HAN ; Xiao-Qun PAN ; Ping LIN ; Routley VIRGINIA ; JoanOzanne-Smith
Chinese Journal of Epidemiology 2009;30(5):459-461
Objective To find out the situation of seat belt wearing among drivers and passengers in Nanjing after the National Road Traffic Safety Law and its Ordinance had been issued for 3 years, in Jiangsu province. Methods Situation on seat belt wearing among drivers and front passengers on four types of vehicles including taxi, cars (below 8 seats), vans and pickups was studied during different time blocks at 4 sites in the city. Results A total number of 35 256 vehicles, their drivers and another 15 772 passengers sitting in the front, were observed. The prevalence rates of seat belt wearing, not wearing and pretend wearing among drivers were 49.9%, 44.1%, and 4.6% respectively while among front-seat passengers were 9.1% and 90.9% respectively. There were significantly declining trends in seat belt wearing among both drivers and front-seat passengers during the 3-years of observation, after adjustment by sex, types of vehicles and other factors (P<0.01). Risk of not wearing among front seat passengers was higher for those 'not-wearing' drivers (OR=8.10, P<0.01). Conclusion Neither the current law nor ordinance in Jiangsu province gives detailed regulations on seat belt wearing, which hampers the effective enforcement.
6.Design of medical UV lamp control system based on development board
Jian-Ning DU ; Dong-Sheng LU ; Cong JIN ; Zi-Han LIN ; Ran WANG ; Min ZHANG
Chinese Medical Equipment Journal 2023;44(12):114-117
Objective To design a medical ultraviolet lamp control system based on a development board to achieve remote control and usage time recording of medical UV lamps.Methods The system was composed of a WeMos D1 mini development board,a 1-way relay module,an OLED display,a RCWL-0516 microwave radar sensor and an ACS712 current monitoring module,which had its control program written by integrated development environment(Arduino IDE).Results Tests proved the system developed functioned well in timed disinfection,manual disinfection and system timing;the ACS712 current monitoring module could detect the lamp failure and current failure;the system could immediately turn off the ultraviolet lamp when persons entered the space to be disinfected;the medical staff could be prompted to replace the lamp when it's about to run out of expiration date.Conclusion The medical UV lamp control system developed gains advantages in easy operation and low cost,and realizes remote control and usage time recording of medical UV lamps.
7.Curative effect of decitabine combined with cytokine-induced killer cells in two elderly patients with acute myeloid leukemia.
Cheng CHANG ; Bo YANG ; Lin ZHANG ; Hong-Li ZHU ; Xue-Chun LU ; Bo GUO ; Li-Li CAI ; Wei-Dong HAN ; Yao WANG ; Hui FAN ; Su-Xia LI ; Yang LIU ; Yang YANG ; Bing ZHAI ; Hai-Hong RAN ; Jie LIN ; Feng ZHANG
Journal of Experimental Hematology 2013;21(1):62-67
This study was aimed to evaluate the effectiveness and safety of low methylation drug decitabine combined with autologous cytokine induced killer cells (CIK) to treat the elderly patients with acute myeloid leukemia (AML). Two AML patients aged over 80 years old were diagnosed and treated in our department from 2006 to 2012; both company with MDS history, and one case was M4-type, another case was M6-type according to FAB classification. The changes in lymphocyte subsets, hematologic response, transfusion frequency, leukemic gene expression, obtaining CR or PR, quality of life and survival time of the patients with different treatment regimen (decitabine alone; CIK alone; decitabine combined with CIK) were systematically observed. The results showed that therapy of decitabine combined with CIK cells could reduce bone marrow suppression extent, decrease the frequency and volume of blood transfusion, and prolong the duration of partial remission, compared with the single use of CIK cell infusion and single use of decitabine treatment. Meanwhile, the kinds of expressed genes associated with leukemia decreased and the survival time was prolonged obviously. The patients' life quality significantly improved. It is concluded that decitabine combined with CIK for treatment of elderly patients with AML is safe and effective.
Aged, 80 and over
;
Azacitidine
;
analogs & derivatives
;
therapeutic use
;
Cytokine-Induced Killer Cells
;
Humans
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Immunotherapy, Adoptive
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Leukemia, Myeloid, Acute
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drug therapy
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therapy
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Male
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Treatment Outcome
8.Development of a Nutrition Questionnaire and Guidelines for the Korea National Health Screening Program for Infants and Children.
Jin Soo MOON ; Jae Young KIM ; Soo Hee CHANG ; Choi Kwang HAE ; Hye Ran YANG ; Jeong Kee SEO ; Jae Sung KO ; Kyung Dan CHOI ; Jeong Wan SEO ; Hee Jung CHUNG ; Baik Lin EUN ; Seung Sik HWANG ; Soon Ryu SEO ; Han Suk KIM
Korean Journal of Pediatric Gastroenterology and Nutrition 2008;11(1):42-55
PURPOSE: A new health policy, referred to as the National Health Screening Program for Infants and Children, was launched in November 2007 by the Ministry of Health and Welfare and National Health Insurance Corporation in Korea. We have developed a nutrition-counseling program that was incorporated into this project. METHODS: We reviewed the nutritional guidelines published by The Korean Pediatric Society and internationally well-known screening programs such as Bright Future in the United States. We also reviewed the recent Korean national surveys on nutritional issues, including the Korea National Health and Nutrition Examination Survey (KNHANES) and the 2005 National Survey of Physical Body and Blood Pressure in Children and Adolescents. The development of questions, pamphlets, computer programs, and manuals for doctors was carried out after several meetings of researchers and governmental officers. RESULTS: We summarized the key nutritional issues according to age, including breastfeeding in infants, healthier complementary feeding, and prevention of iron deficiency anemia, establishment of healthier diets, as well as dietary prevention of overweight children with an emphasis on physical exercise. We have constructed a new Korean nutrition questionnaire and an anticipatory guidance program based on the primary care schedule of visits at 4, 9, 18, 30, and 60 months of age. Five to eight questions were asked at each visit and age-matched pamphlets for parents and guidelines for doctors were provided. CONCLUSION: We developed a nutrition-counseling program based on recent scientific evidence for Korean infants and children. Further research on this national program for screening the nutritional problems in detail and setting the therapeutic approaches may help identify areas of success as well as those that need further attention.
Adolescent
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Anemia, Iron-Deficiency
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Appointments and Schedules
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Blood Pressure
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Breast Feeding
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Child
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Counseling
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Diet
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Exercise
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Health Policy
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Humans
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Infant
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Infant Nutritional Physiological Phenomena
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Korea
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Mass Screening
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National Health Programs
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Nutrition Surveys
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Obesity
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Overweight
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Pamphlets
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Parents
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Primary Health Care
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Surveys and Questionnaires
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Software
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United States
9.Curative efficiency of rituximab combined with autologous cytokine induced killer cells on aged patient with orbital diffuse large B cell lymphoma.
Su-Xia LI ; Hong-Li ZHU ; Bo GUO ; Xue-Chun LU ; Hui FAN ; Jie LIN ; Bo YANG ; Yang LIU ; Yang YANG ; Hai-Hong RAN ; Bing ZHAI ; Wei-Dong HAN ; Yao WANG ; Shan-Qian YAO
Journal of Experimental Hematology 2012;20(5):1117-1121
The aim of this study was to observe the curative effects and safety of autologous cytokine induced killer (CIK) cells in treatment of aged patients with orbital diffuse large B cell lymphoma after rituximab therapy. The patient was given rituximab three times with low dose COP chemotherapy one time when he was diagnosed with orbital diffuse large B cell lymphoma. Two months later, the patient began to receive five cycles CIK cells infusion. One course of therapy was defined as follows: about (2-3)×10(9) of CIK cells (survival rate > 95%) was transfused twice and then rhIL-2 (1 MU daily) was subcutaneously administered for 10 consecutive days. Efficacy and adverse effect was observed during or after CIK cells infusion. The results showed that the peripheral blood mononuclear cells of the patient could be cultured and expanded into CIK cells. The majority of CIK cells was positive for CD3 and CD8 after culture. The CD3(+)CD56(+) cells markedly increased after culture. After two cycles of CIK cell infusion, the orbital lymphoma and possible involvement of the kidney disappeared. The patient obtained complete remission after five cycles of CIK cells infusion. The side effects of CIK cell treatment were minor. It is concluded that CIK cell infusion may prevent recurrence, prolong progression-free survival and improve quality of life after rituximab (alone or with chemotherapy) for aged patients with orbital diffuse large B-cell lymphoma.
Aged, 80 and over
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Antibodies, Monoclonal, Murine-Derived
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therapeutic use
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Combined Modality Therapy
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Cytokine-Induced Killer Cells
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cytology
;
transplantation
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Humans
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Lymphoma, Large B-Cell, Diffuse
;
therapy
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Male
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Orbital Neoplasms
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therapy
;
Rituximab
10.Comparison of sensitivity of audiological tests to identify otitis media with effusion in newborn infants.
Li-hui HUANG ; Xiao-ran MA ; Shuo WANG ; Jun-fang XIAN ; Ling-yan MO ; Hui LIU ; Xiang-rong TANG ; Yi-lin YANG ; Xiao-qing TANG ; Lian-sheng GUO ; De-min HAN
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2008;43(12):886-890
OBJECTIVETo investigate the sensitive factors which were used in routine audiological tests to find out otitis media with effusion (OME) in newborn infants.
METHODSSubjects of this study were 48 infants, including 31 males and 17 females, who failed in the universal newborn hearing screening. The age ranged from 1.5 to 12 months with the average age of 4.3 months. All subjects accepted temporal bone CT and routine audiological assessments, including air-conduction and bone-conduction auditory brainstem response (ABR), 40 Hz-auditory event related potential (40 Hz-AERP), distortion-product otoacoustic emission (DPOAE), acoustic reflex, tympanometries using 226 Hz and 1000 Hz probe tone. Nine factors were statistically analyzed using Kappa test, Univariate chi(2) test and multivariate condition Logistic stepwise regression analysis, which included the results of acoustic immittance, the air-conduction and bone-conduction ABR thresholds, the difference between air-conduction and bone-conduction ABR thresholds, the latency of ABR wave I, duration between ABR wave I and V, 40 Hz-AERP thresholds, amplitudes and thresholds of DPOAE, and acoustic reflex thresholds (ART).
RESULTSSeventy-seven ears were diagnosed with OME, and 19 ears were normal. CT scan of temporal bone was set as a comparative standard. Kappa test indicated that the results of tympanometry with 1000 Hz probe tone (Kappa = 0.745, P < 0.001), the air-conduction ABR threshold (Kappa = 0.453, P < 0.001), the latency of ABR wave I (Kappa = 0.430, P < 0.001), the threshold of 40 Hz-AERP (Kappa = 0.582, P < 0.001), and DPOAE (Kappa = 0.495, P < 0.001) had agreement with the results of temporal bone CT on evaluating the function of middle ear. Univariate analysis indicated that sensitive factors of middle ear function in newborn infants were tympanometry with 1000 Hz probe tone (P < 0.001), ART (P < 0.001), the air-conduction ABR threshold (P < 0.001), the difference between air-conduction and bone-conduction ABR thresholds (P < 0.001), the latency of ABR wave I (P < 0.001), the threshold of 40 Hz-AERP (P < 0.001) and DPOAE (P < 0.001). And multivariate conditional Logistic stepwise regression model showed that tympanometry with 1000 Hz probe tone (P < 0.001) and 40 Hz-AERP threshold (P = 0.004) can be substituted into Logistic stepwise regression equation.
CONCLUSIONSTympanometry with 1000 Hz probe tone and are sensitive factors to find out OME in newborn infants. The air conduction ABR threshold, ABR wave I latency, 40 Hz-AERP threshold and DPOAE could reflect the middle ear function of newborn infants effectively.
Acoustic Impedance Tests ; methods ; Ear, Middle ; physiopathology ; Evoked Potentials, Auditory ; Female ; Humans ; Infant ; Male ; Otitis Media with Effusion ; diagnostic imaging ; physiopathology ; Sensitivity and Specificity ; Temporal Bone ; diagnostic imaging ; Tomography, X-Ray Computed