1. Explant combined with trypsin-digestion for culture of endothelial cells from infantile hemangiomas
Academic Journal of Second Military Medical University 2010;30(2):147-150
Objective: To explore a novel method for cultivation of the endothelial cells(EC) from infantile hemangiomas (IH) in vitro and observe the biological character of cultured endothelial cells. Methods: Fresh operative specimens were obtained from infantile hemangiomas. The endothelial cells of IH were cultured by explant combined with trypsin-digestion technique. Immunohistochemical staining of EnVision method was carried out to identify the cultured cells. The purity of endothelial cell was examined by flow cytometry analysis of FITC-CD34. The biologic characters of endothelial cells were observed under inverted phase contrast microscope. Results: Endothelial cells were successfully cultured from 8 IH specimens of all the 14 explants. The morphology of cultured endothelial cells included two types:polygonal cells and fusifourm cells. The cultured cells were homogenously positive for EC markers-vWF or CD34, indicating their EC origin. Tube formation was found in endothelial cells of 1H. The proportion of CD34+ cells was 76.28% in cultured endothelial cells as detected by flow cytometry analysis. Conclusion: ECs can be successfully isolated and cultured from infantile hemangiomas by explant combined with trypsin-digestion technique; the cultured cells have some characters of endothelial cells.
2. Expression of Fas and Fas ligand in endothelial cells of infantile hemangioma
Academic Journal of Second Military Medical University 2010;29(11):1296-1299
Objective: To examine the expression of Fas and Fas ligand(FasL) in cultured infantile hemangioma endothelial cells (IHECs). Methods: IHECs were isolated and cultured by explant combined with trypsin-digested techniques. Then the expression of Fas and FasL on cultured endothelial cells (ECs) were detected by flow cytometry (FCM) and fluorescent quantitative polymerase chain reaction (FQ-PCR) 2-ΔΔCT method,and the results in ECs were compared with those in Jurkat cells (positive control) and Human umbilical vein endothelial cells(HUVECs,negative control). Results: The positive rate of Fas in cultured IHECs was (90.97±2.36)% as detected by FCM,similar to that in the Jurkat cells ([93.87±1.64]%,P>0.05) and significantly higher than that in the HUVECs (P<0.01). Fas mRNA expression in the cultured IHECs was 1.260±0.721 as detected by FQ-PCR,and was 1.448±0.059 in Jurkat cells,with their ratio being 0.87 and with on significant difference found between the 2 group(P>0.05); the ratio of IHECs to HUVECs (0.354±0.170) was 3.56 and there was significant difference between the 2 groups (P<0.01). The expression of FasL mRNA in cultured IHECs,Jurkat cells and HUVECs was too low as detected by FCM and FQ-RCR to have any biological significance. Conclusion: Fas mRNA is highly expressed in cultured IHECs and FasL is rarely expressed in IHECs, which indicates that apoptosis of IHECs is associated with high expression of Fas in IHECs.
3. Clinical management of hemangiomas and vascular malformations: An analysis of 381 cases
Academic Journal of Second Military Medical University 2010;29(3):311-315
Objective: To discuss the treatment for hemangiomas and vascular malformations. Methods: A retrospective study was conducted on 381 patients with hemangiomas and vascular malformations (January of 1998 to June of 2006). The patients consisted of 167 males and 214 females, with an age range from 1 year to 78 years (median age 16 years). According to the biological classification system introduced by Mulliken in 1982, 98 patients had hemangiomas, 38 had capillary malformations,203 had venous malformations, 27 had arteriovenous malformations and 15 had lymphatic malformations. In all patients with hemangiomas,71 patients were treated with intralesional steroids, 20 patients underwent surgical excision, 7 patients were carefully monitored by doctors. In 38 patients with capillary malformations, 27 underwent sclerosant, others were subjected to laser therapy or surgery. In 203 patients with venous malformation, 136 received sclerotherapy, 21 received surgical excision, and 11 underwent sclerotherapy conjunction plus surgical excision. Twenty-seven patients with arteriovenous malformations received sclerotherapy,interventional therapy,or sclerotherapy conjunction plus surgical excision. Fifteen patients with lymphatic malformations received sclerotherapy conjunction plus surgical excision. Results: All patients were followed-up for 0.5-6 years. Follow-up result showed that 199 patients were cured, 125 were greatly improved,and 46 were partly improved; 10 patients showed no obvious improvement; and 1 patient died. Conclusion: Correct diagnosis and the classification, along with correct treatment strategy, are the keys to increase the treatment outcome and decrease complication of patients with hemangiomas and vascular malformations.
4. Changes of local skin temperature of patients with congenital arteriovenous malformations and its clinical significance
Academic Journal of Second Military Medical University 2006;27(11):1232-1234
Objective: To investigate the changes of local skin temperature in congenital arteriovenous malformations (CAVM) patients before and after treatment, and to assess the relationship between the changes of skin temperature and the outcomes of surgical treatment. Methods: From Sept. 2002 to Apr. 2006, skin temperatures around the lesions were measured in 22 CAVM patients with WMY-01 digital thermometer before and after surgical treatments, and the opposite side or adjacent skins were taken as controls. Furthermore, changes of flow signals and blood flow velocities in the lesions were measured by color Doppler ultrasonography pre- and post-operatively. Results: Skin temperatures of local lesions decreased from (35.58 ±0.68)°C pre-operatively to (34.76±0.63)°C post-operatively(P<0.01); however, the post-operative skin temperature of the lesions was still higher than that of normal skin(34.23±0.83)°C (P<0.05). Post-operatively, color Doppler flow image showed 100% occlusion of blood flow in 1 patient, more than 66.7% occlusion in 14 patients, more than 33.3% but less than 66.7% occlusion in 6 patients, and no occlusion in 1 patient; there was significant difference between those pre- and post-operation(P<0.01). The average blood flow velocity decreased from (80.86±6.97) cm/s pre-operatively to (50.72±5.85) cm/s post-operatively (P<0.01). Conclusion: A significant reduction of surface temperature in CAVMS lesions can be achieved by surgical treatment, and the temperature decrease of the lesion may be used as an indicator for the evaluation of treatment outcomes.
5.Protective Effects of Tempol on Ultraviolet-AI Irradiated Human Dermal Fibroblasts
Shuxian YAN ; Yue HU ; Hui DENG ; Xinyu HONG ; Yu XU ; Kanghuang LIAO
Chinese Journal of Dermatology 2003;0(07):-
Objective To determine the effects of Tempol, one of the nitroxides, in the presence of ultraviolet-AI (UVA1, 340 nm -400 nm) on superoxide enzyme (SOD) activity, lipid peroxidation, and expression of collagen Ⅰ, collagen Ⅲ and matrix metalloproteinases (MMP)-1, MMP-3 in human dermal fi broblasts in vitro. Methods Fibroblasts were irradiated by a single exposure to UVA1 and at the same time incubated with or without Tempol, and detected 24 h later. SOD activity and lipid peroxidation, as shown by accumulation of malondialdehyde (MDA), were detected by biochemical assay. Expression of collagen Ⅰ, collagen Ⅲ (protein levels) and MMP-1, MMP-3 (mRNA level) was examined by ELISA and semi-quantitative RT-PCR, respectively. Results A dose of 15 J/cm2 UVA1 significantly inhibited SOD activity and collagen Ⅰ , collagen Ⅲ protein levels, increased MDA level and stimulated MMP-1, MMP-3 mRNA expression (P
6.Efficacy of low-dose interferon therapy for treating chronic hepatitis C patients who cannot tolerate standard treatment.
Zhen-huan CAO ; Ya-li LIU ; Yan-hong ZHENG ; Shu-dan JI ; Jun-li WANG ; Jian-hua LIAO ; Cun-yuan ZHANG ; Xin-yue CHEN
Chinese Journal of Hepatology 2012;20(1):20-24
OBJECTIVETo investigate the therapeutic efficacy of interferon (IFN) therapy and risk of long-term administration for chronic hepatitis C (CHC) patients who cannot tolerate the standard treatment.
METHODSForty-six CHC patients who had proven intolerant to standard treatments were treated with low-dose IFN (non-pegylated IFN: 60 to 300MIU QOD, or pegylated IFN: 50 to 90 mug/w) plus ribavirin (RBV; 0.6g to 0.9 g/d) for 72 weeks.
RESULTSForty-three (93.5%) of the patients were able to tolerate the long-term treatment with low-dose IFN plus RBV. Only three patients experienced severe side effects (low white blood cell and platelet counts) that required treatment withdrawal. The virology response rates over treatment time were: rapid virologic response (RVR): 10.9%; early virus response (EVR): 30.4%; 24 week virologic response: 45.7%; and, 48 week virologic response: 47.8%. B-sonographic imaging revealed that three patients experienced improved liver morphology through the treatment course. The patients who achieved RVR, EVR, or 24 weeks virologic response also attained higher 48 week virologic response. The 24 week virologic response had the strongest predictive value of good prognosis.
CONCLUSIONSOur study demonstrated that long-term treatment with low-dose interferon plus ribavirin is effective for patients who are otherwise intolerant to standard treatment. In these patients, low-dose IFN plus RBV can obtain a high virologic response rate at 48 week. Furthermore, the 24 week virologic response is sufficiently predictive of treatment success. As with any treatment regimen, it is important for healthcare workers to monitor the disease status and potential side effects throughout the course of therapy.
Adult ; Antiviral Agents ; administration & dosage ; therapeutic use ; Female ; Hepacivirus ; Hepatitis C, Chronic ; drug therapy ; virology ; Humans ; Interferons ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Treatment Outcome
7.Effects of buyang huanwu decoction on the sarcoplasmic reticulum calcium uptake in abdominal aortic constriction induced myocardial hypertrophic rats.
Yan-Pin GU ; Yue-Ling LIAO ; Chen ZHANG ; Wei GUO ; Hong-Chang WEI ; Rong LU
Chinese Journal of Integrated Traditional and Western Medicine 2013;33(5):627-631
UNLABELLEDobjective: To investigate effects of buyang huanwu decoction (BYHWD) on the rats' myocardial hypertrophic model induced by abdominal aortic constriction, and to clarify the molecular regulatory mechanisms for sarcoplasmic reticulum calcium uptake.
METHODSHypertrophic myocardium rat model was induced by abdominal aorta constriction (AAC). Four weeks after modeling, rats were randomly divided into the sham-operation group (Group S), the AAC model group (Group M), the Enalapril group (Group E), and the BYHWD treatment group (Group BYHWD), respectively. The left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), + dp/dtmax,-dp/dtmax, cardiac output (CO), heart mass index (HMI), and left ventricular mass index (LVMI) were observed in each group after 12-week medication. The serum contents of the atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) were detected using ELISA. The SERCA2a activity, the ex- pressions of SERCA2a, phospholamban (PLN), and PLN phosphorylation were observed finally.
RESULTSCompared with Group S, LVSP and LVEDP significantly increased,-dp/dtmax and CO obviously decreased, the myocardial tissue was obviously thickened, the serum contents of ANP and BNP increased, the activity and expression of SERCA2a decreased, the SERCA2a/PLN ratio and PLN phosphorylation degree decreased in Group M (all P <0.05). Compared with Group M, LVEDP obviously decreased, -dp/dtmax and CO obviously increased, the hypertrophy myocardial tissue was obviously lessened, the serum contents of ANP and BNP decreased, the activity of SERCA2a increased, the relative expression contents of SERCA2a, Ser16, and Thrl7 were elevated in Group BYHWD (all P <0.05). BYHWD had significant roles in elevating the SERCA2a/PLN ratio and PLN phosphorylation degree (P <0.05).
CONCLUSIONBYHWD could significantly improve hemodynamics of heart failure rats, elevate CO, lessen cardiac hypertrophy, and improve the capabilities for sarcoplasmic reticulum calcium uptake.
Animals ; Aortic Aneurysm, Abdominal ; metabolism ; pathology ; Calcium ; metabolism ; Constriction, Pathologic ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Hypertrophy, Left Ventricular ; metabolism ; pathology ; Male ; Myocardium ; pathology ; Rats ; Rats, Wistar ; Sarcoplasmic Reticulum ; metabolism
8.Study on the unintended pregnancy among married women of child-bearing age living in Qingshan district, Wuhan
Yu-Liang ZOU ; Si-Lan LIAO ; Hua YANG ; Ru-Cheng CHEN ; Dan-Qin HUANG ; Shi-Yue LI ; Hong YAN
Chinese Journal of Epidemiology 2012;33(3):291-295
Objective To investigate the prevalence of unintended pregnancy (UP) and exploring the risk factors of UP for married women of child-bearing age from Qingshan district,Wuhan.Methods A cross-sectional study was adopted in this study.Cluster sampling method was used with 3256 women recruited,in 2010.Information on history and risks related to social-demographic factors of UP were collected,using a self-administered questionnaire.Results Of the 3256 participants,over half of them (53.8%) reorted ever having had the history of UP and 9.1% reported UP in the past year.Rate of UP in the past year for different age cohorts (18-30,31-40,41-49 years) were 31.8%,10.5% and 1.8% respectively.The most frequently reported reason for UP across all the age cohorts was "Did not use any contraceptive methods",with proportions on the reason that reported by women at 18-30,31-40 and 41-49 year-olds,were 69.7%,51.1% and 42.4% respectively.The second frequently reported reasons for UP were "Failure of traditional contraception" for younger cohort ( 18-30 years:13.0% ) and "IUD dropped or pregnancy with IUD" for older-age cohorts (23.4% at 31-40 year-olds and 37.0% at the 41-49 year-oplds).The most frequently cited reason for "Did not use any contraceptive methods" was "Believe we were lucky so far,not to get pregnant" (59.6%).The risk factors of UP were being at older age,experiencing sex debut at younger age and got married at younger age.Conclusion The prevalence of lifetime UP history was high among women at child-bearing age from Qingshan district,Wuhan.Reproductive health services and interventions should be taken according to the needs from different age cohorts of women.Younger cohort of women should receive more attention.
9.Simultaneous determination of four constituents in Hydnocarpus anthelmintica by HPLC
Xue MA ; Huan ZOU ; Xu SUN ; ting Yue LI ; gao Shang LIAO ; hong Jun LIU ; jun Yong LI
Chinese Traditional Patent Medicine 2017;39(11):2325-2328
AIM To establish an HPLC method for the simultaneous content determination of four constituents in Hydnocarpus anthelmintica Pierre.METHODS The analysis of 60% ethanol extract from H.anthelmintica was carried out on a 35 ℃ thermostatic Diamonsil C18column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of 80% acetonitrile-0.1% phosphoric acid flowing at 1.0 mL/min in a gradient manner,and the detection wavelength was set at 347 nm.RESULTS Luteolin,hydnocarpusol,chrysoeriol and hydnocarpin showed good linear relationships within the ranges of 0.0523 6-1.047 μg (r =0.999 9),0.011 24-0.224 8 μg (r =0.999 9),0.029 46-0.589 2 μg (r =0.999 9) and 0.130 5-2.609 μg (r =0.999 9),whose average recoveries were 102.45% (RSD=1.9%),98.66% (RSD=1.8%),97.60% (RSD=1.6%) and 97.88% (RSD=1.4%),respectively.CONCLUSION This simple,accurate and sensitive method can be used for the quality control of H.anthelmintica.
10.Effect of tumor necrosis factor-it on the expression of ATP binding cassette transporter A1 in THP-1 macrophage foam ceils
Hong-Yu PENG ; Zhi-Jian CHEN ; Yu-Hua LIAO ; Yan-Fu WANG ; Chun-Li MEI ; Bing YUE ; He-Ping GUO ; Min WANG
Chinese Journal of Geriatrics 2000;0(04):-
Objective To investigate the influence of tumor necrosis factor-?(TNF-?) on ATP binding cassette transporter A1(ABCA1)in THP-1 macrophage foam cells, and the intervention effect of nuclear factor-?B(NF-?B) inhibitor TPCK on the TNF-?, so as to determine the role of TNF-?/ NF-?B in cellular cholesterol efflux. Methods Foam cells were transformed from THP1 cells. The correlation of cellular cholesterol efflux from foam cells with different concentrations and time stimulated by TNF-? were estimated. Subsequently foam cells were treated with TNF-? at satulated concentration(10.0 ng/ml ), TPCK(10?mol/L),or TPCK(10?mol/L) pretreated for 60 min before TNF-? stimulation. ABCA1 gene expression was analyzed by RT-PCR. ABCA1 protein level was detected by Western blot. Results TNF-? decreased cellular cholesterol efflux of foam cells in concentration-and time-dependent manner. 10 ng/ml of TNF-? down-regulated the levels of both ABCA1 mRNA and protein expressions in time-dependent manner. TPCK was observed to efficiently block the suppressive effect of TNF-? on ABCA1. Conclusions TNF-? decreases cellular cholesterol efflux mainly through the down-regulation of ABCA1. TPCK, an inhibitor of NF-?B activation, is observed to partly block the suppressive effect of TNF-? on ABCA1, suggesting a mechanism involving NF-?B signal transduction. TNF-?/NF-?B might play a critical role in the progression of atherosclerosis by decreasing cellular cholesterol efflux from foam cells.