OBJECTIVETo investigate the effects of Dragon' s blood extract on proliferation and secret extracellular matrix function of fibroblasts in vitro.

METHODSDragon' s blood was extracted by chloroform, acetoacetic ester, alcohol. Human fibroblast were cultured in vitro in media containing gradient dilutions of Dragon' s blood extracts (0.002, 0.02, 0.2, 2, 20 mg/ml) , which was followed by cell proliferation assessed with MTT assay on 0, 12, 24, 36, 48, 60, 72 h. Under the optimal concentration, the cell growth curves were drawn and the flow cytometry (FCM) was used to determine the changes of cell cycle. On 0, 12, 24, 36, 48, 60, 72 h, the concentration of hyaluronic acid in the supernatant of fibroblast culture was measured by radioimmunoassay.

RESULTS0.2-2 mg/ml Dragon' s blood extracts enhanced the proliferation of fibroblasts in a dose-dependent manner. 2 mg/ml was the optimal dilution of Dragon's blood extract, and it increased the ratio of S cells in cell cycle [(25.80 ± 3.10)%] than control group [(7.50 ± 0.70)%, P < 0.01]. From 12 h to 72 h, in 2 mg/ml Dragon's blood group, concentration of Hyaluronic acid secreted by fibroblasts gradually increased, but were less than control (P < 0.01).

CONCLUSIONSDragon's blood acetoacetic ester extract improved the proliferation of cultured human fibroblasts in vitro, might be beneficial to promote wound healing.

Cell Cycle ; Cell Proliferation ; drug effects ; Culture Media ; chemistry ; Dose-Response Relationship, Drug ; Extracellular Matrix ; Fibroblasts ; cytology ; drug effects ; secretion ; Flow Cytometry ; Humans ; Hyaluronic Acid ; analysis ; secretion ; Plant Extracts ; pharmacology ; Resins, Plant ; Time Factors

BACKGROUND:Dragon’s blood is the main ingredient of traditional medicine prescription for promoting granulation, which has been used in clinical treatment of a variety of refractory wounds and achieved the exact effects. But the Dragon’s blood effect on colagen secretion from normal fibroblasts has not been reported. OBJECTIVE: To investigate the effects of Dragon’s blood extract on the proliferation and secret function of fibroblastsin vitro. METHODS: Dragon’s blood was extracted by extracts chloroform, acetoacetic ester, and alcohol in turn. Normal human fibroblasts were respectively cultured in Dragon’s blood extracts of chloroform, acetoacetic ester, and alcohol, DMEM containing 1% dimethyl sulfoxide, and normal culture medium. Then, the fibroblasts were cultured in vitro in different media containing gradient dilutions of Dragon’s blood extracts (0.002, 0.02, 0.2, 2, 20 g/L), which was folowed by cel proliferation determination assessed with MTT assay. Under the optimal concentration, the cel growth curves were drawn and the flow cytometry was used to determine the changes of cel cycle. The concentration of procolagen type III in the supernatant of the fibroblast culture systems was measured by radioimmunoassay. RESULTS AND CONCLUSION:0.2 g/L-2 g/L dilution of Dragon’s blood extracted by acetoacetic ester enhanced the proliferation of fibroblasts in a dose-dependent manner. The 2 g/L was the optimal dilution of Dragon’s blood extracted by acetoacetic ester, and it increased the ratio of S cels in cel cycle than control group and decreased procolagen type III. These findings indicate that Dragon’s blood acetoacetic ester extract can improve the proliferation of cultured human fibroblastsin vitro, and decrease the secretion of procolagen type III of fibroblasts, and it can be beneficial to improve wound healing and inhibit hypertrophic scar.

Objective:To investigate the supply status of dementia care services in pension agency in Guang zhou,thus to put forward suggestions for improving the service quality of the long-term care for dementia.Methods:Using questionnaire research and individual interview,an interview survey was conducted among managers,dementia and nursing assistant in ten pension agencies in Guangzhou.The data was analyzed by SPSS16.0 statistical method.Results:The dementia had been increasingly paid attention by the pension agencies,but there were still considerable defects in the aspect of the zone set up and service supply;the professional quality of nursing personnel in pension agencies had steadily increased,but shorthanded situation was more obvious.Conclusions:It suggests that we should provide specialized nursing level and the classification standard of care requirements for dementia elders;strengthen the training of mental health worker to enhance the level of medical care and supply ability of nursing assistant;allocate the mental health resources rationally;explore the “combination of medical treatment and endowment” service model which meets the need of dementia elders.

Objective To investigate the clinical efficacy of fluoroscopy-guided intestinal adhesion lysis, as a new non-surgical method, in treating incomplete adhesive small intestinal obstruction in order to improve the therapeutic results of adhesive intestinal obstruction. Methods A total of 93 patients with incomplete adhesive small intestinal obstruction were enrolled in this study. The patients were divided into study group (n=49) and control group (n=44). Fluoroscopy-guided intestinal adhesion lysis together with restoration of inter-intestinal loop enterocele was carried out for the patients of the study group , while traditional conservative surgical therapy was employed for the patients of the control group. The study group was comparable with the control group in patients’ age, gender, medical history, disease course, X-ray findings, etc. Results Of the 49 cases in the study group, complete cure was obtained in 40 with a cure rate of 81.6%. The mean hospitalization day was 0.3 day, and the average operation time was 3.25 hours. Among the 44 patients in the control group, complete cure was obtained in 37 with a cure rate of 84.1%. The mean hospitalization day was 7.6 days, and the average therapeutic time was 183.26 hours. Conclusion For the treatment of incomplete adhesive small intestinal obstruction , the therapeutic efficacy of fluoroscopy-guided intestinal adhesion lysis together with restoration of inter-intestinal loop enterocele is better than that of traditional conservative surgical therapy.

Objective To explore the clinical value of gonadotropin-releasing hormone analogue (GnRH-a) by analyzing the efficiency of GnRH-a treatment for moderate and severe endometriosis after laparoscopic conservative operations.Methods Two hundred and sixty-five females who accepted laparoscopic conservative operation were enrolled in this study,and they were confirmed by pathology for ovarian endometriosis cyst,in phase Ⅲ andⅣ,47 patients had fertility desire.According to whether use the drug and auxiliary species the postoperative cases were divided into 5 groups:102 cases without endocrinal therapy(untreated group),GnRH-a group of 3 months in 64 cases,GnRH-a group of 6 months in 31 cases;gestrinone group of 3 months in 38 cases,and gestrinone group of 6 months in 30 cases.All patients were followed up for 2 years,observed drug clinical effects,remission rate,improvement rate,recurrence rate,pregnancy outcomes and side effects.Results The remission rate in GnRH-a group of 6 months and gestrinone group of 6 months was significantly higher than that in untreated group [83.9％ (26/31),73.3％ (22/30) vs.52.9％(54/102)],the remission rate in GnRH-a group of 6 months was significantly higher than that GnRH-a group of 3 months [64.1％ (41/64)],there was significant difference (P ＜ 0.05).The recurrence rate in GnRH-a group of 3 months,GnRH-a group of 6 months,gestrinone group of 6 months was significantly lower than that in untreated group [12.5％ (8/64),9.7％ (3/31),10.0％ (3/30) vs.27.5％ (28/102)] (P ＜ 0.05).The recurrence rate in gestrinone group of 6 months was significantly lower than that in gestrinone group of 3 months (P ＜ 0.05).The recurrence time of GnRH-a group of 3 months,GnRH-a group of 6 months,gestrinone group of 6 months was significantly longer than that in untreated group [(14.05 ± 1.97),(16.76 ± 1.53),(16.12 ±2.15)months vs.(12.85 ± 1.80)months] (P ＜0.05).The recurrence time of gestrinone group of 3 months and untreated group had no significant difference(P ＞ 0.05).The pregnancy rate of GnRH-a group and gestrinone group was higher than that in untreated group,but there was no significant difference (P ＞ 0.05).The rate of menopausal symptom in GnRH-a group was higher than that in gestrinone group [50.5％ (48/95) vs.16.2％ (11/68)].The rate of abnormal bleeding in vagina,acne and side-effect of aminotransferase in GnRH-a group were significantly lower than those in gestrinone group [1.1％(1/95) vs.7.4％(5/68),0 vs.8.8％(6/68),0 vs.5.9％(4/68)](P＜ 0.05).Conclusions Laparoscopic conservative operations combined with GnRH-a on treatment of moderate and severe ovarian endometriosis can significantly improve remission rate,decrease recurrence rate.GnRH-a or gestrinone for 6 months,the pregnancy rate increases.In addition the side reactions of the two medicine have statistic difference,which of GnRH-a is lower.

Objective To clarify the effectiveness of QS Nd ∶ YAG laser on melasma in Chinese patients.Methods A group (n=45) of patients previously diagnosed as facial melasma were treated with QS Nd ∶ YAG laser at approximately 1 week intervals.Eleven to twenty treatments were per formed for each patient.The treatment efficacy of QS Nd ∶ YAG laser was evaluated by patients and doctors,respectively,at various time point such as before treatment,5 weeks and 10 weeks after treatment and end of the treatment.The skin melanin index and the transepidermal water loss were e valuated by the skin multifunction tester at different time point,respectively.Results Compared with untreated,the melanin index of melasma areas decreased significantly after the laser treatment for 5 weeks,10 weeks and the end of treatment.But there was no significant difference in the melanin index of normal skin areas with or without laser treatment.Compared with untreated,the transpidermal water loss of melasma areas increased significantly after the laser treatment for 5 weeks,10 weeks and end of treatment.The patients' skin became smooth,delicate,pores shrink and more flexible.In 45 melasma patients treated by QS Nd ∶ YAG laser,8 cases basically cured (17.78 ％),25 cases were markedly effective (55.56 ％),12 cases improved (24.44 ％) and only 1 case was uneffective (2.22 ％).Theoverall effective rate was 73.33 ％.Conclnsions QSNd∶ YAG laser is a useful treatment modality for Chinese women who have melasma with precise efficacy,less side effects and high safety.

Objective To study the relationship of levels of circulating endothelial microparticles (EMP62E, EMP31) and high-sensitivity C-reactive protein (hs-CRP) with severity of chronic left heart failure in elderly patients. Methods According to New York Heart Association (NYHA) class and left ventricular ejection fraction (LVEF), the healthy subjects and the patients were divided into five groups: control group [LVEF: (63.97±4.65)%], classⅠ group [LVEF: (42.67±2.06)%], classⅡ group [LVEF: (34.26±3.17)%], class Ⅲ group [LVEF: (29.05±1.07)%] and class Ⅳ group[ LVEF:(25.17±1.42)%] . The levels of circulating EMP62E, EMP31 and hs-CRP of the patients and healthy subjects were measured by flow cytometry and nephelometry immunoassay, respectively. Results There were significantly differences in EMP62E, EMP31 and hs-CRP between class Ⅳ group and classⅠ group P＜0.01) EMP62E [(1092.7 ± 102.8) counts/μl vs. (291.0±21.9) counts/μl], EMP31 [(1596.1±46.3) counts/μl vs. (477.8±40.3) counts/μl] and hs-CRP [(14.74±0.07) mg/L vs. (4.86 ± 0.09) mg/L]. The levels of circulating EMP62E, EMP31 and hs-CRP were gradually elevated significantly along with the increased severity of chronic left heart failure in elderly. Conclusions The upregulation of circulating EMP31, EMP62E and hs-CRP may contribute to the development of chronic heart failure in elderly.

Objective To observe the effect of changes in CD34+ cell level on various degrees of chronic heart failure (CHF) in the elderly. Methods The enrolled patients were divided into four CHF groups according to the New York Heare Association(NYHA) functional class: NYHA class I (n=23), Ⅱ (n=27), Ⅲ (n = 20) and IV group (n= 16) , and there were 41 healthy controls over the same period. The levels of peripheral blood CD34+ cells were measured, and the tumor necrosis factor-α (TNF-α). its soluble receptors (sTNFR-1 and sTNFR-2) and vascular endothelial growth factor (VEGF) were also measured. Results The levels of CD34+ cells were elevated in the early CHF and depressed in the advanced CHF in elderly patients. The levels of CD34+ cells were (0. 6± 0.2) 109/L in control group, (2.4±0. 4) 109/L in NYHA class I group, (1.9±0.2)×109/L in NYHA class Ⅱ group, (1.3±0.1)×109/L in NYHA class Ⅲ group and (0.5±0.2)×109/L in NYHA class Ⅳ group, respectively (all P<0.01).And TNF-α, sTNFR-1, sTNFR-2 and VEGF were increased in severe chronic left ventricular failure CNYHA class IV vs. I group:TNF-α: (61.4± 15.7) ng/L vs. (28.4±10.8)ng/L; sTNFR-1: (2820.9±1282.8)ng/L vs. (690.8±62.7) ng/L; sTNFR-2: (4113.1±1102.2) ng/L vs. (740.8± 112.3)ng/L; VEGF: (996.3±487.1)ng/L vs. (423. 3±147. 9)ng/L, all P<0. 013. Conclusions The changes of CD34+ cell level may predict various degrees of chronic heart failure in elderly patients.

Objective To investigate the influence of gene transfection antiangiogenesis on microvessel and relative cytokine of hypertrophic scar of rabbits' ear.Methods The hypertrophic scar of rabbtis' ear was reproduced.On the 10th day after epithelization,Ad-METH-1 was injected into tissue of scar.30 days later,the microvessel of scar-tissue was detected by microcirculation microscope.Meanwhile.H&E and immunohistochemical stains were performed.Then the results were analyzed.Results 30 days after Ad-METH-1 injection.in experimental groups,the microvascular count of scar tissue was 12.38±2.56,the percentage of VEGF positive cells was 17.64%,and the percentage of bFGF positive cell was 18.24%:while in the control groups,the microvascular count of scar tissue was 48.12±6.46.the percentage of VEGF positive cell was 31.34%.and the percentage of bFGF positive eell was 28.26%.Results revealed that the count of microvessel of scar tissue in the experimental groups was lower than that in the control groups,between which there was the difference in statistics(P＜0.01).and that the percentage of VEGF and bFGF positive cells of scar tissue in the expenmental groups were lower than that in the control groups.between which there was the difference in statistics(P＜0.05).Conclusion Ad-METH-1 has marked inhibitory effects on scar tissue hyperplasia of rabbits' ear,angiogenesis and expression of VEGF and bFGF.Using antiangiogenesis therapy at the early phase could inhibit the formation of hypertrophic scar.Gene transfection antiangiogenesis therapy could bid fair to become an effective method to prevent and treat hypertrophic scar.

Objective To analyze the alterations of serum protein in ESCC,compare alterations of serum protein with and without LM. Methods Serum samples were collected from 64 ESCC patients before operation and 60 cases with gender and age-matched healthy controls,special serum protein or peptide spectra was determined by SELDI-TOF-MS measurement after treating the sample onto weak cation exchange (WCX2) protein chip for each case. The serum protein profiles were compared by Biomarker Wizard Software between the ESCC patients and healthy controls, and among ESCC patients stratified according to gender, age, location of tumor, size of tumor, infiltration and with or without LM. Results (1)120 protein peaks were detected at the molecular range of 0 to 50000 in comparing of ESCC patients and healthy controls. 31 significantly different peaks were found between ESCC patients and healthy controls (P <0.05), 10 peaks were selected(P<0.01). (2) One significantly different protein peak (m/z 4174) was detected between T1 and T3, T4 (P<0.05). (3) There were three significantly different protein peaks (m/z 3970,4174 and 4277) between with LM and without LM (P<0.05).The peak (m/z 4174) was shared by two groups above. (4) No significant different protein was found when patients stratified according to gender, age, location of tumor and size of tumor. Conclusion Significant difference exists in serum proteins between ESCC patients and healthy controls. There are statistical difference exists in serum proteins between T1 and T3, T4, with LM and without LM. This difference is less than between ESCC patients and healthy controls. Some commonness is existed in serum protein fingerprint for patients with serious infiltration and with LM.