1.Expression of miRNA-1247-3P in acute respiratory distress syndrome cell model and its function
Dongliang CHENG ; Yuan LIANG ; Yanchen CHEN ; Di QING ; Changsong SHI
The Journal of Practical Medicine 2015;(17):2802-2804
Objective To detect the expression pattern of microRNA in A549 cells treated by lipopolysaccharide, study the expression of miRNA-1247-3P in A549 cells under LPS treatment and explore the possible mechanism of miRNA-1247-3P in A549 cells under LPS treatment. Methods A549 cells were divided into experimental and control groups. Immunocytochemical method and RT-PCR were used to detect the changes of SP-A and SP-C. The expression of miRNAs were detected using miRNAs array in different groups. The key miR-1247-3P was collected to detect the changes of miR-1247-3P in all groups using quantitative real-time polymerase chain reaction. Results Compared with control group, the expressions of SP-A and SP-C were significantly decreased in the experimental groups (P < 0.05). MiRNA array showed that 31 miRNAs were up-regulated and 3 miRNAs were down-regulated. Compared with control group, the expression of miR-1247-3P was significantly increased in the experimental groups (P < 0.05). Conclusion The increased expression of miR-1247-3P may play an important role in the pathogenesis of ARDS.
2.Three different fixation instruments for tibial fractures:a biomechanical comparison
Jun-Wu HUANG ; Xiang-Yang WANG ; Cheng-Di SHI ;
Chinese Journal of Orthopaedic Trauma 2004;0(07):-
Objective To compare the biomechanical performance of three different fixation instruments for tibial fractures.Methods Fourteen fresh tibial specimens were made into models of oblique fracture.Seven models were fixed with unilateral axial dynamic fixation(UADF),and seven with limited contact dynamic compression plate(LC-DCP).After biomechanical tests had been done for the UADF group,an extra screw was used at the oblique fracture site to reinforce the fixation with extra limited internal fixation.Each model was tested for its hiomechanical performance in resisting compression,bending and rotation.Results The performance of UADF was significantly poorer than that of LC-DCP and UADF with limited internal fixation in anti-compression,an- ti-bending and anti-rotation(P<0.05).There was no significant difference in the rigidity between LC-DCP and UADF with limited internal fixation(P>0.05).Conclusion UADF with extra limited internal fixation is a recommendable method for tihial fractures because it cart provide the same effective fixation as LC-DCP does.
3.Diagnostic Value of Lymphoscintigraphy in Chylous Ascites in Children
chu-gang, LI ; hong-liang, FU ; de-di, XU ; cheng-ren, SHI
Journal of Applied Clinical Pediatrics 2004;0(12):-
Objective To evaluate the value of lymphoscintigraphy in diagnosis of chylous ascites in children.Methods Lymphoscintigraphy was done in 6 cases,computed tomography(CT) was done in 4 cases,X-ray exam was done 42 times.And their video repore were compared.Results Lymphoscintigraphy was done in 6 cases,5 cases′ results were positive which diagnosed chylous ascites,and their leaking positions were also found.Conclusion Lymphoscintigraphy has the qualitative and orientational effect on diagnosis of children with chylous ascites.
4.The expression and purificiation of human Annexin Ⅴ overexpressed in E. coli and the detection of apoptosis
Chunhui DI ; Qun SHI ; Yingmei ZHANG ; Cheng ZHAO ; Yingcheng ZHONG ; Dawu HUANG ; Dalong MA
Journal of Peking University(Health Sciences) 2001;33(2):167-169
Objective: in order to provide rapid and reliable method. Methods: Encoded Annexin Ⅴ cDNA was amplifyed from U937 cDNA libary by PCR and then subcloned into E coli expression vector. MS2-Annexin Ⅴ fusion protein could be overexpressed in E coli. The MS2 bacteria protein could be removed by thrombin digestion.The mature Annexin Ⅴ was obtained by ion exchange chromatography and the FITC labled Annexin Ⅴ could be used in the detection of apoptosis. Results:Up to 37% of the total bacterial proteins was rhAnnexin Ⅴ as showed by SDS-PAGE. The purification of Annexin Ⅴ is over 99%. The FITC labled Annexin Ⅴ could efficiently detect apoptosis. Conclusion: We successfully established the technique procedure of obtaining a large quantity of Annexin Ⅴ and provided the basic routine for popularizing the detection of apoptosis' with high effciency.
5.Effect of kebimin decoction in treating allergic rhinitis and on blood levels of nitric oxide and superoxide dismutase.
Shi-fu ZHANG ; Li-lan CHENG ; Di-nan HUANG
Chinese Journal of Integrated Traditional and Western Medicine 2004;24(2):118-120
OBJECTIVETo observe the therapeutic effect of Kebimin decoction (KD) on allergic rhinitis (AR) and its effect on blood levels of nitric oxide (NO) and superoxide dismutase (SOD) activity.
METHODSEighty-two AR patients were randomly divided into two groups, the treated group and the control group, 41 in each group. To the treated group, KD was given one dose per day for 10 days as one therapeutic course and 1-3 courses were given successively. The control group was treated with Xinfang Rhinitis capsule for 30 days. Blood levels were determined and compared before and after treatment.
RESULTSThe total effective rate in the treated group was 93%, which was better than that in the control group (51%), the difference was significant (chi 2 = 17.704, P < 0.01). Serum level of NO was higher and that of SOD activity was lower in the AR patients than that in healthy persons (P < 0.01), KD could significantly lower the former and increase the latter (P < 0.01).
CONCLUSIONThe therapeutic effect of KD in treating AR was significant, its mechanism might be related with the lowering of NO and increasing of SOD activity in serum, as well as the scavenging of oxygen free radical.
Adolescent ; Adult ; Child ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Free Radical Scavengers ; therapeutic use ; Humans ; Male ; Middle Aged ; Nitric Oxide ; blood ; Phytotherapy ; Rhinitis, Allergic, Perennial ; blood ; drug therapy ; Superoxide Dismutase ; blood
6.Bidirectional effect of MnSOD overexpression on the proliferation of esophageal cancer cells in vitro.
Guo-Gui SUN ; Ya-Di WANG ; Cheng-Lin LI ; Yun-Jie CHENG ; Shao-Wu JING ; Qing LIU ; Shi-Jie WANG
Chinese Journal of Oncology 2011;33(8):583-589
OBJECTIVETo construct a recombinant lentiviral vector for manganese superoxide dismutase (MnSOD) gene expression, and observe its effect on the proliferation of esophageal cancer cells in vitro.
METHODSChemical methods were employed for synthesis of the MnSOD cDNA sequence sections, along with the attB sites. Target gene fragment was constructed on the pMD-18T vector, and the recombinant plasmid pDONR221 was obtained after BP recombination reaction. Sequencing was followed by LR recombination reaction between the plasmid and DEST to obtain the lentiviral vector, which worked with helper plasmid for co-transfection of human embryonic kidney epithelial cells (293T cells). Amplification was done to determine its titer, and both transfection and selection procedures were made to get two stable transfected esophageal cancer TE-1 cell lines with medium MnSOD expression (TE-1Mm cells) and high MnSOD expression (TE-1Mh cell), and empty vector cell (TE-1Mn cells). Reverse transcription polymerase chine reaction (RT-PCR), immunofluorescence, immunocytochemistry and Western blot were used to detect the target gene with respect to its expression in the TE-1 cells. Additionally, colorimetric 3-[4,5-dimethy thiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, agar colony formation assay, annexin V-FITC/PI staining and flow cytometry experiments were also conducted as to observe the influence of the medium and high MnSOD overexpressions on the proliferation of esophageal cancer cells.
RESULTSRT-PCR indicated that the transfected TE-1 cells showed positive MnSOD expression at different levels. Immunofluorescence, immunocytochemistry and Western blot suggested that TE-1Mm cells and TE-1Mh cells had MnSOD protein expression at different levels. MTT assay indicated that TE-1Mm cells had a significantly decreased survival rate compared with that of the two control cells (TE-1 cells and TE-1Mn cells), and TE-1 Mh cells had an significantly increased survival rate (P<0.05). The colony formation ability of TE-1Mm cells was (23.0 +/- 2.7)%, and that of TE-1Mh cells was (45.3 +/- 4.5)%, significantly different form the (34.7 +/- 4.2)% in TE-1 cells and (33.7 +/- 4.7)% in TE-1Mn cells (P<0.05). Annexin V-FITC/PI double staining experiment of the stably transfected cells cultured for 48 h showed that the early apoptosis rate in TE-1Mm cells was (10.6 +/- 1.0)%, significantly higher than (2.6 +/- 0.2)% in the TE-1 cells, (2.5 +/- 0.6)% in the empty vector cells and (1.0 +/- 0.1)% in the TE-1Mh cels (P<0.05). The fluorescence index (FI) of mitochondrial apoptosis of TE-1Mm cells was 0.948 +/- 0.019, significantly lower than that of TE-1 cell (1.000 +/- 0.022) and empty vector The fluorescence index of TE-1Mn cells (0.997 +/- 0.023) and TE-1 cells (1.000 +/- 0.022) were significant different from that of 0.948 +/- 0.019 in TE-1Mm cells and 1.076 +/- 0.022 in TE-1Mh cells, indicating a significant difference of mitochondrial apoptosis between the cell groups. FCM results indicated that the ROS fluorescence index of TE-1Mm cells was 0.859 +/- 0.040, that of TE-1Mh cells was 0.763 +/- 0.039, significantly lower than that of TE-1 cells (1.000 +/- 0. 042) and empty vector cells (1.002 +/- 0.047) (P<0.05).
CONCLUSIONSStably transfected cell lines with MnSOD expression have been successfully established. MnSOD overexpression shows bidirectional effect on the proliferation of esophageal cancer cells.
Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Esophageal Neoplasms ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; HEK293 Cells ; Humans ; Lentivirus ; genetics ; Mitochondria ; pathology ; Plasmids ; RNA, Messenger ; metabolism ; Reactive Oxygen Species ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Superoxide Dismutase ; genetics ; metabolism ; Transfection
7."Medium and Long-term Plan for the Prevention and Control of Chronic Non-communicable Diseases in Shanghai(2018-2030)" in the perspective of public health
Qun-di YANG ; Ping-ping BAO ; Yang ZHENG ; Min-na CHENG ; Ye RUAN ; Chun-fang WANG ; Yan SHI
Shanghai Journal of Preventive Medicine 2020;32(6):487-
The "Medium and Long-term Plan for the Prevention and Control of Chronic Non-communicable Diseases in Shanghai (2018-2030)" was officially released in August 2018.From the perspective of public health, this paper analyzes the background of the plan from the epidemic situation, response and challenges Shanghai City is facing, expounds the comprehensive prevention and control system of chronic diseases including four functional systems, and explains the key preventive and control measures on the different stages of chronic diseases, comparing the evaluation indicators with those of the national plan.This paper will help to better understand the new blueprint for the prevention and control of chronic diseases in Shanghai in the next ten years.
8.Medium-term follow-up of laparoscopic surgery on radical gastrectomy with D2 dissection for gastric cancer
Yu MING ; Xue DI-XIN ; Shi ZHENG-CHAO ; Chen CHENG-LIANG ; He XIN-WEI ; Chen JI-XIAN ; Ma XIAO-HONG
China Journal of Endoscopy 2017;23(11):68-73
Objective To explore the feasibility and clinical outcome of laparoscopy-assisted gastrectomy with D2 dissection by comparing the efficacy of open surgery on radical gastrectomy with D2 dissection for gastric cancer patients. Methods The patients with gastric cancer from October 2008 to August 2013 were divided into two groups according to the different surgical methods. Among them, 175 patients underwent laparoscopic surgery (laparoscopy-assisted surgery group, Group L), and 163 patients underwent laparotomy surgery (open surgery group, Group O). The number of lymph nodes dissected, postoperative recovery, complications, mortality and survival rate of the two groups were compared and analyzed. Results There was no significant difference in the number of lymph nodes resect between the two groups [Group L and Group O: (26.3 ± 13.9) vs (26.8 ± 10.2), t = -0.40, P = 0.684]. Compared with open surgery, the laparoscopy-assisted surgery showed significantly less intraoperative blood loss and quicker recovery of gastrointestinal function. The postoperative hospitalization time of laparoscopic group was less than that of laparotomy group, the difference was statistically significant (P < 0.05). No significant difference was found in 3-year survival rate between the two groups (Group L vs Group O: 92.00% vs 92.63%, P = 0.262). Conclusions Compared to open surgery, laparoscopic gastrectomy is safer and has quicker postoperative recovery. There is no statistical difference in the number of resect lymph nodes between lapamscopic and open gastrectomy.
9.A standard protocol for detection of EGFR mutations in cytologic specimens.
Zheng WANG ; Xiaonan WU ; Yuankai SHI ; Xiaohong HAN ; Gang CHENG ; Lin LI ; Li ZHANG ; Yuhui ZHANG ; Xinlin MU ; Guangqing ZHU ; Zaiwen FAN ; Li YANG ; Jing DI ; Xinrui JIA ; Dongge LIU
Chinese Journal of Oncology 2014;36(7):516-521
OBJECTIVEThe aim of this study was to establish a standard protocol for detection of EGFR mutations in cytologic specimens.
METHODS287 cytologic samples were collected from the patients who were suspected of having lung cancer at six hospitals in Beijing. A detection protocol for EGFR mutations was designed. Two comparative experiments were carried out for the coincidence in EGFR mutation rates between direct sequencing (Seq) and amplification refractory mutation system (ARMS) methods, and between 40 matched cytologic samples with formaldehyde-fixed paraffin embedded (FFPE) cytologic blocks and cytospin slides.
RESULTSTumor cells were found in 236 out of 287 cases (82.2%, 236/287) . Among them, there were 31 cases (13.1%, 31/236) of low tumor cell content samples and 205 cases (86.9%, 205/236) of high tumor cell content samples. 180 cases in the high tumor cell content samples (87.8%, 180/205) were diagnosed to be consistent with NSCLC. 25 out of 194 cases were ruled out or indefinite to be diagnosed as NSCLC by immunohistochemistry. By direct sequencing, the mutation rate of EGFR was 27.8% (50/180) in NSCLC samples and 28.2% (50/177) in adenocarcinoma samples (high tumor content samples) . By ARMS, the mutation rate of EGFR was 45.6% (82/180) in NSCLC samples and 46.3% (82/177) in adenocarcinoma samples (high tumor content samples). The EGFR mutation rate in low tumor content samples was 38.7% (12/31) , there was no significant difference in EGFR mutation rates between the groups of low tumor cell content samples and high tumor cell content samples (P = 0.12). The concordance rate of EGFR mutation rates was 100% between scraping tumor cells from slides samples and from FFEP blocks in the 40 matched samples. Forty-eight out of 180 definitive NSCLC patients received Gefitinib therapy. The FPS was 12 months in the gefitinib-treated ARMS⁺ group and 2 months in the ARMS⁻ group (P < 0.001), and the OS was 19 months in the gefitinib-treated ARMS⁺ group and 7 months in the ARMS⁻ group (P = 0.003), but no significant differences were found in the efficacy (PFS and OS) of Gefitinib between Seq⁺ and Seq⁻ groups (P = 0.227, P = 0.510, respectively), and Seq⁺/ARMS⁺ and Seq⁻/ARMS⁺ groups (P = 0.354, P = 0.334, respectively).
CONCLUSIONSThe detection protocol for EGFR mutations in cytological specimens introduced in this study is tested to be reliable and feasible. Pathological evaluation and immunohistochemistry are important in the detection procedure of EGFR mutations in cytologic specimens. High sensitivity methods should be selected for detection of EGFR mutations in cytologic samples.
Adenocarcinoma ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; Humans ; Lung Neoplasms ; diagnosis ; epidemiology ; metabolism ; Mutation ; Mutation Rate ; Polymerase Chain Reaction ; Receptor, Epidermal Growth Factor ; genetics ; metabolism
10.Intestinal preparation of compound polyethylene glycol electrolyte powder combined with Magnesium Sulfate in constipation patients
Xiao-Fen MENG ; Di ZHANG ; Yi YANG ; Hua LI ; Ya-Ya TIAN ; Cheng-Ying SHI ; Ping-Min QU
China Journal of Endoscopy 2018;24(4):38-41
Objective To investigate the effect of compound polyethylene glycol electrolyte powder (SF-PEG) and Magnesium Sulfate (MgSO4) on intestinal tract preparation in patients with constipation. Methods 135 cases of constipation patients who underwent colonoscopy were selected. They were divided into A, B and C groups, each with 45 cases.Group A and group B received oral SF-PEG and 219.2 g (2 000 ml) at 5:00 to 7:00. Group A take 50% MgSO450 ml at 9:00, and then took 250 ml warm water.Group B received the same dose of MgSO4 at 7:00. The two groups were examined by colonoscopy at 11:00. Group C received oral compound SF-PEG 219.2 g (2 000 ml) at 10:00 to 12:00, and then took 50% MgSO450 ml at 14:00, received colonoscopy at 16:00. According to the Boston bowel preparation scale (BBPS) score and parallel intraluminal bubble score. Determine the duration of bowel preparation process,and tolerance and adverse effects were recorded during bowel preparation. Results All patients completed bowel preparation and underwent a colonoscopy successfully. The scores of group A in BBPS were significantly higher than those in group B and group C (P < 0.05). In group A and group B, the score of parallel intraluminal bubble was lower than C, which was statistically significant (P < 0.05). The intestinal preparation time of A and B was less than that of group C, which was statistically significant (P < 0.05), and the tolerance of patients was higher in group A and group B than that in group C (P < 0.05). In adverse reactions, group A and group B were lower than those in group C (P < 0.05). Conclusions When taking the time (5:00 to 7:00), intermittent polyethylene glycol electrolyte powder and Magnesium Sulfate can shorten the bowel preparation time and improve the quality of bowel preparation in patients with constipation.