1.Homing of bone marrow hemopoietic stem cells in infarct area following myocardial infarction
Chinese Journal of Tissue Engineering Research 2009;13(27):5252-5256
BACKGROUND: Studies have shown that after bone marrow stem cell transplantation, there are juvenile cells with the episode of myocardial cells around the infarct areas, which was recognized as deriving from bone marrow stem cells.OBJECTIVE: To explore the homing situation of bone marrow stem cells by autologous mobilization around the infarct areas aftar myocardial infarction.DESIGN, TIME AND SETTING: The cytological in vitro study was performed at the China-Japan Clinical Pathology Center, Daliar Medical University from February 2005 to April 2006.MATERIALS: A total of 30 myocardial samples from the autopsy at the Department of Pathology and Forensic Medicine, Dalian Medical University between 2000 and 2005. Fifteen normal samples dyed from the mechanical injuries or craniocerebral injury,without abnormity following autopsy. Fifteen samples in the myocardial infarction group suffered from coronary atherosclerosis and coronary tube stenosis, and died within 10 days after myocardial infarction.METHODS: The autopsy was performed within 48 hours after the death and the tissues were obtained from antedor wall, lateral wall and posterior wall of the left ventricle, and made into tissue sections. We checked the expression of CD34, CD133 and transforming growth factor-β, in the normal area, infarct area and around the infarct area by immunohistochemistry.MAIN OUTCOME MEASURES: The expression of CD34, CD133 and transforming growth factor-β in different areas and the relationship between them.RESULTS: The positive expression of CD34 in infarct area was significantly stronger than that around the infarct area and in normal area (P < 0.01). The positive expression of transforming growth factor-β in infarct area was significantly stronger than that around the infarct area (P < 0.01), and the expression of transforming growth factor-β showed negative reactJon in normal area. The expression of CD34, CD133 and transforming growth factor-β, in infarct areas and around the infarct area was closely related (P < 0.01).CONCLUSION: There are the homing of bone marrow hemopoietic stem cells in infarct area and around the infarct ares after myocardial infarction. The increase in transforming growth factor-β in infarct area provides the possibility for the differentiation of the homing stem cells, the following angiogenesis and the regeneration of cardiomyocytes.
2.Evaluation of nasal continuous positive airway pressure on prethrombotic state in obstructive sleep apnea syndrome
Chinese Journal of Practical Internal Medicine 2006;0(13):-
Objective To estimate the effects of nasal continuous positive airway pressure(nCPAP)therapy on patients with OSAS in prethrombotic state.Methods One group of 30 patients with OSAS who were diagnosed from Sep.2003 to Sep.2005,one healthy control group of 30 cases.Blood samples were taken from people in the supine position in the morning immediately after overnight polysomnography and nCPAP theraphy(only in OSAS group).Results Compared with the normal controls,the level of the molecular marker GMP-140 increased in the blood of patients with OSAS,the levels of APTT,AT-Ⅲ decreased,and the level of t-PA decreased as well.The level of PAI increased.Compared with the healthy controls,datum indicated statistical differences between the two groups.Datum of the patients with OSAS were improved after nCPAP therapy:AHI decreased from 63.21?23.23 to 0.56?0.84;the nocturnal lowest of SaO_2 increased from 0.68?0.07 to 0.90?0.02;GMP-140 and PAI decreased to (17.07?7.97)% and (95.83?5.91)% respectively;AT-Ⅲ and t-PA increased to(95.20?6.67)% and (5.45?1.61)?g respectively.Conclusion Prethrombotic state is present in patients with OSAS.nCPAP therapy can decrease OSAS-related thrombotic complications.
3.KAP-1 knock down by shRNA imparts sphere-forming activity of Panc-1 cells
Jianxin JIANG ; Lei ZHAN ; Yaozhen PAN ; Chengyi SUN
Chinese Journal of Hepatobiliary Surgery 2013;19(7):520-525
Objective To investigate whether KAP-1 could induce CSC-self renewal in pancreatic cancer cell lines.Methods KAP-1 expression was examined in 14 cases of pancreatic cancer with immunohistochemistry.KAP-1 shRNA amplified by PCR was inserted into pGC-LV in vector,and then identified by restriction endonuclease digestion and nucleotide sequencing.The lentiviral vector pGC-shRNA-KAP-1 was co-transfected with pHelper 1.0 and pHelper 2.0 packaging plasmids into HEK 293T cells,and the lentivirus was collected and virus titer was measured.The expressions of KAP-1 and vimentin were detected by Western blot and RT-qPCR when human pancreatic cancer cell line Panc-1 was infected by the lentivirus.Sphere forming assay was conducted to assess the capacity of CSC or CSC-like cell self-renewal in this study.Results The KAP-1 expression level in cancerous tissues on immunohistochemistry was significantly higher than in the corresponding normal tissues (P=0.002).After infected by lentivirus,the expressions of KAP-1 and vimentin were knocked down,which could be detected by Wesren blot and RT-qPCR.Compared to Panc-1-GFP (NC) cells,the outcomes suggested that knocking down the expression of KAP-1 could decrease the formation of pancreatospheres,which further suggested the capacity of CSC-self-renewal in primary and secondary pancreatospheres of Panc-1-shRNA-KAP-1 and NC cells.Conclusions KAP-1 expression in pancreatic cancer tissues has been identified.The lentiviral vector for shRNAs targeting KAP-1 was constructed successfully.The formation of pancreatospheres decreased by knocking down the KAP 1 gene.KAP-1 is involved in the regulation of CSC phenotype.The mechanism is probably related to the upregulated expression of the EMT marker vimentin.
4.Effects of KAP-1 in promoting the epithelial-mesenchymal transition of pancreatic cancer cell line Capan-2
Chengyi SUN ; Jianxin JIANG ; Yaozhen PAN ; Lei ZHAN
Chinese Journal of Digestive Surgery 2013;12(8):586-591
Objective To investigate the effects of KAP-1 in promoting the epithelial-mesenchymal transition (EMT) of pancreatic cancer cell line Capan-2.Methods The lentiviral vector of LV-plenti-GFP-KAP-1 was constructed.Capan-2 cells were divided into the experimental group (cells transfected by lentiviral vector of LVplenti-GFP-KAP-1),negative control group (cells transfected by empty vector) and blank control group 1 (cells cultured in 1640 medium plus 10% fetal calf serum).Capan-2 cells in the experimental group were subdivided into the miR-100-5p inhibitor transfection group (cells transfected with miR-100-5p inhibitor),empty vector control group (cells transfected with microRNAs inhibitor),blank control group 2 (cells cultured in 1640 medium plus 10% fetal calf serum).The lentivirus was identified by double endonuclease restriction and sequencing,and the virus titer was detected.The morphological changes of the cells were observed after transfecting lentiviral vector of LV-plenti-GFP-KAP-1 to the Capan-2 cells.The expressions of KAP-1,genes of EMT proteins and mRNA of miR-100-5p were detected by real-time quantitative polymerase chain reaction.The protein expressions of KAP-1,EMT proteins in all the groups were detected by Western blot.The measurement data were presented by mean ± standard deviation,and were analyzed using the analysis of variance.Results The lentiviral vector of LV-plentiGFP-KAP-1 was successfully constructed,and the virus titer was 2 × 108 TU/ml.Compared with the control group,the mesenchymal transition of the Capan-2 cells was detected in the experimental group after transfecting the Capan-2 cells with lentiviral vector of LV-plenti-GFP-KAP-1 for 48 hours.The relative mRNA expressions of KAP-1,N-cadherin,vimentin,E-cadherin,miR-100-5p were 1.77 ± 0.83,2.62 ± 0.71,2.50 ± 0.21,7.20 ± 1.17 and 1.81 ±0.40 in the experimental group,5.03 ±0.29,5.07 ±1.53,3.83 ±0.57,7.83 ±0.78,7.01 ± 0.96 in the negative control group,5.13 ± 1.14,5.81 ± 1.49,4.92 ± 0.90,3.07 ± 0.36,6.87 ± 0.35 in the blank control group 1,with significant difference among the 3 groups (F =5.99,7.62,7.88,6.62,4.64,P <0.05).The relative mRNA expression of KAP-1 in the miR-100-5p inhibitor transfection group,empty vector control group,blank control group 2 were 1.56 ± 0.42,4.89 ± 0.61,5.20 ± 0.38,with significant difference among the 3 groups (F =5.14,P < 0.05).The relative mRNA expressions of vimentin in the miR-100-5p inhibitor transfection group,empty vector control group,blank control group 2 were 3.10 ± 1.37,3.44 ± 0.94,3.08 ±1.16,with no significant difference among the 3 groups (F =0.49,P > 0.05).The results of western blot showed that the relative protein expressions of KAP-1,N-cadherin,vimentin,E-cadherin were 2.77 ± 1.99,1.31 ±0.38,4.25 ± 0.63,0.62 ± 0.06 in the experimental group,0.83 ± 0.46,0.41 ± 0.37,1.03 ± 0.33,1.17 ± 0.45 in the negative control group,0.71 ± 0.26,0.08 ± 0.04,1.37 ± 0.92,3.04 ± 0.65 in the blank control group 1,with significant difference among the 3 groups (F =5.54,4.68,3.19,8.18,P < 0.05).The relative protein expression of KAP-1 in the miR-100-5p inhibitor transfection group,empty vector control group,blank control group 2 were 2.27 ±0.71,0.56 ±0.43,0.61 ±0.39,with significant difference among the 3 groups (F =4.81,P <0.05).The relative protein expressions of vimentin in the miR-100-5p inhibitor transfection group,empty vector control group,blank control group 2 were 3.19 ± 0.55,3.93 ± 0.06,3.61 ± 0.73,with no significant difference among the 3 groups (F =0.04,P > 0.05).Conclusion KAP-1 promotes the EMT of Capan-2 cells by specifically down-regulating the miR-100-5p expression.
5.Etiological analysis of asymptomatic hematuria in 431 children and follow-up
Zhicai SUN ; Yuling LIU ; Xiaofen PAN ; Guomo LI ; Xinyan LEI
Journal of Clinical Pediatrics 2015;(9):810-812
ObjectiveTo investigate the etiology and prognosis of asymptomatic hematuria in children.MethodsThe etiological factors, clinical features and prognosis of asymptomatic hematuria were analyzed retrospectively in 431 children from Jan. 2001 to Dec . 2014. ResultsIn 431 children (197 males and 234 females) with asymptomatic hematuria, the mean age of ifrst visit was 5.52±2.77 years (8 months-17 years). Four hundred and twenty-ifve cases had persistent microscopic hematuria and 6 cases had gross hematuria. Three hundred and iffteen cases (73.1%) were glomerular hematuria, among which 286 cases were isolated hematuria, 5 cases were acute glomerulonephritis, 13 cases were minimal change glomerulopathy, 4 cases were IgA nephropathy, 4 cases were mesangial proliferation glomerulonephritis and 3 cases were thin basement membrane nephropathy. One hundred and thirty-six cases (31.5%) were non-glomerular hematuria, among whom 113 cases were left renal vein entrap-ment syndrome, 17 cases were idiopathic hypercalciuria, 4 cases were kidney stone, 1 case was urinary tract infection and 1 case was left kidney absence. The mean follow-up period was 3.05±2.69 years (0.5-13.5 years). One hundred and forty-ifve patients showed the resolution of microscopic hematuria, among whom 110 cases (75.8%) had the resolution in 3 years after the ifrst visit. In 24 cases with family history of hematuria, only 6 cases showed the resolution. At the end of the follow-up, renal function remained stable in all children.ConclusionsThe onset age of asymptomatic hematuria in children varies widely, and most of them are glomerular hematuria. Most children with isolated hematuria show resolution within three years after the ifrst visit. The children with familial hematuria may last longer. The isolated hematuria has good prognosis but needs to be followed up.
6.Characteristics of fall injuries among primary and middle school students in Yinzhou District from 2010 to 2022
SUN Qiang ; ZHAO Lei ; LI Xiaoyong ; PAN Wenjie
Journal of Preventive Medicine 2024;36(3):207-210
Objective:
To investigate the characteristics of fall injuries among primary and middle school students in Yinzhou District, Ningbo City, Zhejiang Province from 2010 to 2022, so as to provide insights into prevention and control of fall injuries among primary and middle school students.
Methods:
Data of 6 to 17 year-old primary and middle school students with initial diagnosis of fall injury from 2010 to 2022 were collected through Injury Subsystem of Zhejiang Provincial Chronic Disease Surveillance Information Management System. The time, place, activity and clinical characteristics of fall injury were analyzed using a descriptive method.
Results:
Totally 7 808 fall injury cases among primary and middle school students were reported in Yinzhou District from 2010 to 2022, accounting for 45.72% of the total injury cases in the same age. There were 5 413 boys and 2 395 girls, with a boy/girl ratio of 2.26∶1. Primary school students were the majority, accounting for 55.10%. The incidence of fall injuries among girls, junior high school students and high school students showed increasing trends from 2012 to 2022 (both P<0.05). The primary peak of fall injury was from September to November, and the secondary peak was from May to July, with 4 510 cases (57.76%). The place of fall injury development mainly included schools (2 680 cases, 34.32%), homes (2 343 cases, 30.01%) and streets/urban areas (2 247 cases, 28.78%). The activities at the time of fall injury mainly included leisure time (3 490 cases, 44.70%), sports (2 861 cases, 36.64%) and school activities (1 094 cases, 14.01%). Soft tissue injury was the main characteristics (6 224 cases, 79.71%). Lower limbs (3 101 cases, 39.72%), head (2 419 cases, 30.98%) and upper limbs (1 974 cases, 25.28%) were the main injury sites. Mild injury was predominant (5 896 cases, 75.51%).
Conclusions
Boys and primary school students are high-risk groups of fall injury in Yinzhou District, schools are high-risk places of fall injury. Schools should be regarded as key intervention places and health education on fall injury prevention should be strengthened.
7.In vitro transdermal delivery of Qingfei Xiaocuo gel based on principal component analysis.
Wei-gao REN ; Lin-xiu PENG ; Fei-fei LEI ; Cheng-xiang SUN ; Jin-huo PAN
China Journal of Chinese Materia Medica 2015;40(2):231-235
The objective of the present study was to establish a method based on principal component analysis (PCA) for the study of transdermal delivery of Chinese medicinal formulae, and to choose the best penetration enhancers for Qingfei Xiaocuo gel depend on this method. Using improved Franz type diffusion cell and excised rat skin in vitro as transdermal barrier, the receptive solution fingerprint was established by HPLC, harvesting the areas of the common peaks in the fingerprint, then the total factor scores of the concentrations at different times were calculated using PCA and were employed instead of the concentrations to compute the cumulative amounts (Q12) and enhancement ratio (ER), the latter of which were considered as the indexes for optimizing penetration enhancers. Compare to the control group, the ER of the other groups increased significantly and furthermore, 2.5% azone with 2.5% menthol manifested the best effect. PCA represent most information in the receptive solution, the method above could choose the best penetration enhancers, it could be a reference for the study of transdermal delivery of Chinese medicinal formulae.
Administration, Cutaneous
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Animals
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Drugs, Chinese Herbal
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analysis
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pharmacokinetics
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Gels
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In Vitro Techniques
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Male
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Medicine, Chinese Traditional
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Mice
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Principal Component Analysis
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Skin
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metabolism
8.Relationship between adipose cytokines and metabolic syndrome in newly diagnosed type 2 diabetes mellitus patients
Qi PAN ; Lixin GUO ; Lei JIANG ; Mingxiao SUN ; Ming LI ; Xiaoping LIU ; Yao WANG
Chinese Journal of Geriatrics 2009;28(1):26-29
Objective To investigate the relationship between serum levels of adiponectin (APN) and retinol binding protein 4 (RBP4) in newly diagnosed type 2 diabetes mellitus (T2DM) patients. Methods 95 newly diagnosed T2DM patients and 55 non-diabetic volunteers from health examination population were studied. The serum concentrations of APN and RBP4 were determined by ELISA. The levels of plasma glucose, lipid profiles and insulin were also measured in fasting samples. Insulin sensitivity was assessed by insulin resistance index (HOMA-IR). The control group and diabetic group were divided into ≥60 years subgroup and <60 years subgroup respectively. According to the guideline designed by Chinese Diabetes Society(CDS)in 2004, the control group and diabetic group were divided into different kinds of abnormal metabolism groups. Results (1)The level of APN was significantly decreased in diabetic group compared with the control group[(7.26±4.69)mg/L vs. (11.93±4.89)mg/L], while the serum level of RBP4 in diabetic group was obviously higher than that in control group[(16.48±7.82) mg/L vs. (10.91±5.26) rag/L]. (2) The prevalence of metabolic syndrome in diabetic group was higher than that in control group (46.3% vs. 7.3%). (3)APN level was decreased with the increasing number of components of MS, and RBP4 level was increased with the increasing number of components of MS. (4) In DM group, APN level was correlated negatively with MS, waist circumference and BMI, and RBP4 level was correlated positively with waist circumference, HOMA-IR, BMI and TG. Conclusions APN and RBP4 levels are significantly associated with metabolic syndrome. Hypoadiponectinemia and higher retinal binding protein 4 may play an important role in the pathogenesis of MS.
9.Cloning and analyzing the activity of the promoter of the MUC1 gene in Panc-1 cells
Yaozhen PAN ; Chengyi SUN ; Lei ZHAN ; Hao ZHANG ; She TIAN ; Hong ZHANG ; Nan JIANG
Chinese Journal of Hepatobiliary Surgery 2014;20(2):137-141
Objective To evaluate the MUC1 promoter's role in driving gene expression in pancreatic cancer and its therapeutic significance.Methods Two plasmids were made.The plasmid pEGFP-MUC1N1 contained MUC1 promoter fragment connected to the pEGFP-N1 vector with the EGFG reporter gene.The pShuttle-MUC1-EGFP plasmid contained MUC1 promoter fragment and EGFP reporter gene connected to pShuttle plasmid.Lipofectamine 2000 was used to transfect the two plasmids into cells of MUC1-positive human pancreatic cell line Panc-1 and MUC1-negative human cervical carcinoma Hela.Fluorescence microscopy and flow cytometry compared the specificity and activity of the MUC1 promoter and CMV promoter.Results Reporter gene EGFP-positive cells 48 hours after transfection with pEGFP-MUC1-N1 and pShuttleMUC1-EGFP plasmid were 69.6% and 63.6% respectively,in Panc-1 cells,and 4.2% and 3.7% respectively,in Hela cells.Conclusions MUC1 promoter can drive reporter gene activity in MUC1-positive tumor cells targeting functional expression.There is potentially a use of targeted therapy in pancreatic cancer at the genetic level.
10.Emergency plastic treatment in children's facial injury
Lei PAN ; Sufan WU ; Sheng YAN ; Hua WU ; Yi SUN ; Da CHEN
Chinese Journal of Medical Aesthetics and Cosmetology 2014;20(3):196-198
Objective To analyze and summarize the features of children's facial wound and to share the emergency treatment experiences.Methods An retrospective study was conducted on the pathogenesis,age distribution,emergency treatment methods and complications in 1407 children (under 15 years of age) during recent 8 years (from January 2004 to December 2011).Results In this study the major facial wound type was simply lacerations (1196/1407,85.0 %),and the most common wound cause was trumbling injury (1154/1407,82.0 %).The T area was the easiest injured,successively,forehead (562/1407,39.9%),chin (211/1407,15.0%),lips (196/1407,13.9%) and nose (95/1407,6.8%).Here were 2 peaks in age distribution which were 2 and 5 years of age,respectively.Pleased outcome would be found while normalized plastic means were taken.After 3 to 6 months following up,there were 5 infection cases,27 cases healed with hypertrophic scars,and 7 cases with traumatic tattoos in wound location.Conclusions The most common wound type is simply laceration.2 and 5 years of age are two of the highest risk stages.It is necessary to act up to plastic surgery principles and take children's particularity into account when emergency treatment,and to do our best to minimize the scar in the first operation time.