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Objective To observe the analgesic effects of mild moxibustion on primary dysmenorrhea(PD)rats with cold and dampness stagnation syndrome and its effect on BDNF/TrkB signaling pathway in hypothalamus;To explore its mechanism for the treatment of PD.Methods A total of 32 Wistar non-pregnant female rats were randomly divided into blank group,model group,Western medicine group and mild moxibustion group,with 8 rats in each group.Except for the blank group,the other groups received estradiol benzoate intraperitoneal injection combined with ice bath treatment + oxytocin intraperitoneal injection to establish PD with cold and dampness stagnation syndrome model.The mild moxibustion group received treatment at"Shenque"and"Guanyuan"from the eighth day of modeling for 10 min,and the Western medicine group was given ibuprofen solution intragastically for 4 days.The latency period of rats twisting was observed and the twisting score was calculated,Western blot and PCR were used to detect the expressions of c-fos,BDNF,TrkB protein and mRNA in hypothalamic tissue.Results Compared with the blank group,the model group showed a shortened latency period and an increased twisting score(P<0.01),the expressions of c-fos,BDNF,TrkB protein and mRNA in hypothalamic tissue increased(P<0.01,P<0.05).Compared with the model group,the mild moxibustion group had a longer latency period and lower twisting score(P<0.01),while the expressions of c-fos,BDNF,TrkB protein and mRNA in hypothalamic tissue increased(P<0.01,P<0.05).Conclusion Mild moxibustion may effectively improve the pain state of PD rats with cold and dampness stagnation syndrome.This mechanism may be related to downregulating c-fos expression,inhibiting BDNF/TrkB signaling pathway activation,thereby inhibiting pain signal transmission,regulating pain occurrence and maintenance.
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Objective:To observe the effects of sparrow-pecking moxibustion at Shenque(CV8)and Guanyuan(CV4)on the writhing reaction and score,the temperature and blood flow perfusion of moxibustion point area and uterus,the serum levels of arginine vasopressin(AVP),prostaglandin(PG)F2α,and thromboxane(TX)B2 in rats with primary dysmenorrhea(PD)due to cold-dampness stagnation,and to explore the possible mechanism of sparrow-pecking moxibustion in treating PD.Methods:Thirty-two healthy non-pregnant female Wistar rats were randomly divided into a normal group,a model group,an ibuprofen group,and a sparrow-pecking moxibustion group,with 8 rats in each group.Except for the normal group,the other three groups were subjected to modeling with cold water bath combined with estradiol benzoate and oxytocin injection.Rats in the sparrow-pecking moxibustion group were treated with sparrow-pecking moxibustion at Shenque(CV8)and Guanyuan(CV4)on the 8th day of modeling,30 min/time,once a day for 3 d;those in the ibuprofen group were treated with 0.8 mL ibuprofen solution(a specification of 125 mg in 10 mL)on the 8th day of modeling,once a day for 3 d;those in the normal group and the model group were given 0.8 mL normal saline,once a day for 3 d.On the 11th day,rats in each group were intraperitoneally injected with oxytocin(2 U/rat),and the writhing incubation period and writhing score in 20 min were observed;the temperature and the blood perfusion of Shenque(CV8),Guanyuan(CV4),and uterus in vivo were detected;the serum levels of AVP,PGF2α,and TXB2 were determined.Results:The writhing incubation period was significantly longer(P<0.01)and the writhing score was significantly lower(P<0.01)in the sparrow-pecking moxibustion group and the ibuprofen group than in the model group;compared with the ibuprofen group,the writhing incubation period was prolonged(P<0.01)and the writhing score was decreased(P<0.01)in the sparrow-pecking moxibustion group;compared with the normal group,the temperature and the blood perfusion of Shenque(CV8),Guanyuan(CV4),and uterus were significantly decreased,while the serum PGF2α,AVP,and TXB2 levels were significantly increased(P<0.01)in the model group;compared with the model group,the temperature and the blood perfusion of Shenque(CV8),Guanyuan(CV4),and uterus were significantly increased,and the serum levels of PGF2α,AVP,and TXB2 were significantly decreased in the ibuprofen group and the sparrow-pecking moxibustion group(P<0.05 or P<0.01);compared with the ibuprofen group,the temperature and the blood perfusion of Shenque(CV8),Guanyuan(CV4),and uterus were significantly increased(P<0.05),the serum AVP and TXB2 levels were significantly decreased(P<0.05),while the serum PGF2α level had no statistical difference in the sparrow-pecking moxibustion group(P>0.05).Conclusion:Sparrow-pecking moxibustion had a remarkable analgesic effect on the rats with PD due to cold-dampness stagnation,and the mechanism may be related to the increased temperature and blood perfusion of the moxibustion point area and uterus,as well as the decreased serum PGF2α,AVP,and TXB2 levels.
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Objective: To observe the effects of ginger-partitioned moxibustion at Shenque (CV8) and Guanyuan (CV4) on the expression levels of endocrine-related molecules and their receptors in rats with primary dysmenorrhea (PD) due to cold-dampness stagnation, thus to explore their analgesic mechanisms. Methods: Thirty-two female Wistar rats were divided into a normal group, a model group, a ginger-partitioned moxibustion group, and a Western medicine group according to the random number table method, with 8 rats in each group. Except for rats in the normal group, all other rats were treated with oxytocin combined with ice-water bath to establish the rat models of PD due to cold-dampness stagnation. After successful modeling, rats in the normal group and the model group did not receive treatment; rats in the ginger-partitioned moxibustion group received treatments with ginger-partitioned moxibustion at Shenque (CV8) and Guanyuan (CV4); rats in the Western medicine group received ibuprofen by intragastric administration. The writhing response of rats was compared among groups, and the serum levels of prostaglandin F2α (PGF2α), estrogen (estradiol, E2), progesterone (P), and the mRNA expression of PGF2α and E2 receptors in the uterine tissues were detected. Results: No writhing behavior was observed in the normal group; compared with the normal group, the serum PGF2α and E2 levels in the model group were increased (P<0.01), while the P level was decreased (P<0.01), and the mRNA expression levels of the uterine PGF2α and E2 receptors were increased (P<0.01, P<0.05). Compared with the model group, the writhing behavior latency was prolonged, and the writhing response score was decreased in the ginger-partitioned moxibustion group and the Western medicine group (P<0.01); the serum PGF2α and E2 levels in the ginger-partitioned moxibustion group and the Western medicine group were decreased, while the P level was increased (P<0.05 or P<0.01); the mRNA expression levels of the uterine PGF2α and E2 receptors in the ginger-partitioned moxibustion group and the Western medicine group were decreased (P<0.05). Compared with the Western medicine group, the ginger-partitioned moxibustion group showed a prolonged writhing behavior latency, reduced writhing response score (P<0.05), and decreased serum E2 level (P<0.05), while no statistical differences in the serum PGF2α and P levels, or the mRNA expression levels of uterine PGF2α and E2 receptors (P>0.05).Conclusion: The analgesic effect of ginger-partitioned moxibustion on PD due to cold-dampness stagnation may be related to regulating the mRNA expression levels of PGF2α and E2 receptors in the uterine tissues.
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Objective To investigate the expression of gastric and intestinal phenotypic markers in Siewert typeⅡand Ⅲ early gastroesophageal junction(GEJ) cancer, and to explore its correlation with clinic-pathological features.Methods From April 2010 to July 2015, 53 cases diagnosed as early GEJ cancer were enrolled.The gastric and intestinal phenotypic markers such as mucin5AC(MUC5AC),mucin6(MUC6),mucin2(MUC2),caudal related homeodomain transcription 2(CDX2) and cluster of differentiation 10(CD10) were detected, and then the patients were divided into gastric type, gastrointestinal type, intestinal type and non-classified type according to the results of immunohistochemical staining.Combined with Siewert classification the clinicopathological features were analyzed.Chi square test or Fisher′s exact test was performed for statistical analysis.Results In the cancer tissues of 47 patients with Siewert type Ⅱand Ⅲ early GEJ cancer, the case numbers of positive expression of MUC5AC,MUC6,MUC2, CDX2 and CD10 were 21(44.7%),19(40.4%),31(66.0%),27(57.4%) and 17(36.2%),respectively;the case numbers of gastric type, gastrointestinal type, intestinal type and non-classified type were 11(23.4%),14(29.8%),21(44.7%) and one(2.1%), respectively.The positive expression rates of MUC5AC and MUC6 in Siewert typeⅡwere 55.9%(19/34) and 50.0%(17/34),which were higher than those of Siewert typeⅢ(2/13), and the positive expression rate of MUC2 was 55.9%(19/34), which was lower than that of Siewert typeⅢ(12/13), and the differences were statistically significant (x2=6.240,4.679 and 4.053;all P<0.05).In Siewert typeⅡ, the proportion of intestinal type was 32.4%(11/34), which was lower than that of Siewert typeⅢ(10/13), and the differences were statistically significant (x2=7.142,P=0.010).In patients with Siewert typeⅡand Ⅲ early cancer, males predominated in intestinal type which were mostly well differentiated type with less submucosal carcinoma.The maximum diameter of tumor was less than those of gastric type and gastrointestinal type.In paracancerous mucosal tissues, the incidences of intestinal metaplasia in gastrointestinal type and intestinal type were 11/14 and 81.0%(17/21), which were higher than that of gastric type (3/11);the incidences of atrophy in gastrointestinal type and intestinal type were 12/14 and 85.7%(18/21),which were higher than that of gastric type (4/11),and the differences were statistically significant (Fisher′s exact test,all P<0.05).Conclusions Siewert typeⅡand Ⅲ early GEJ cancer can directly originated not only from gastric mucosa, but also from gastrointestinal and intestinal mucosa.Atrophy and intestinal metaplasia could exist before cancer genesis.
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Objective To determine the effects and dosage of N?acetylcysteine( NAC) in the im?provement of the visibility in esophagogastroduodenoscopy( EGD) . Methods A prospective randomized con?trolled study was performed on 193 patients scheduled for EGD from November 2014 to July 2015 were ran?domized into five groups using digital table. In group A, 100 mg dimethicone and 2 g NaHCO3 were given. In group B,100 mg dimethicone, 2 g NaHCO3 and 20 000 U pronase were given. Group C received 100 mg dimethicone, 2 g NaHCO3 and 200 mg NAC. Group D received 100 mg dimethicone, 2 g NaHCO3 and 400 mg NAC and group E 100 mg dimethicone, 2 g NaHCO3 and 600 mg NAC.The agents were dissolved in 100 ml water for each patient.Endoscopy was completed by one endoscopist and the score of image visibility assessment was completed by 2 other endoscopists. The 3 endoscopists were unaware of grouping. The total scores, the time of washing, the time of examination and complications were compared and analysed. The total image scores of group A, B, C,D and E were 30?83±3?78, 35?69±2?88, 33?16±3?90, 34?95±3?46 and 36?76±2?91, respectively. Group A was the lowest(P0?05).Images that were scored 3 were the most in group E.The washing times of each group were 38?00±19?10, 17?03±11?44, 15?92±10?81, 15?78 ±10?24 and 15?55±9?69, and the examination times of each group were 13?49±2?49, 9?41±1?86, 9?08± 1?80, 8?73±1?91 and 8?78±1?79 minutes.Group A was the longest in these two indices(P0?05) . Conclusion The preoperative NAC can improve the visibility in EGD. The best dose is 600 mg, whose effects and safety were similar to those of 20 000 U, but yield to less washing time and the examination time in EGD.
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Objective To explore the correlation between serum growth differentiation factor‐15(GDF‐15) level and acute myocardial infarction(AMI) to provide a basis for the prognostic evaluation of AMI .Methods Totally 192 Han patients with AMI (AMI group) and non‐coronary heart disease (NCHD ,NCHD group) diagnosed in Chengde Municipal Central Hospital from Sep‐tember 2013 to January 2015 ,were selected and their clinical data were collected .The biochemical markers and serum GDF‐15 level were detected .Results Comparing the AMI group with the NCHD group ,differences in the patients′age ,smoking ,blood glucose (Glu) ,TC ,TG ,LDL‐C levels had statistical significance (P<0 .05);the serum GDF‐15 level in the AMI group was significantly higher than that in the NCHD ;serum GDF‐15 level was positively correlated with TC ,LDL‐C ,hs‐CRP and Glu in the AMI group . Conclusion The increase of serum GDF‐15 level is obviously correlated AMI ,therefore GDF‐15 can serve as an indicator for moni‐toring myocardial infarction .
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On-off firing patterns, in which repetition of clusters of spikes are interspersed with epochs of subthreshold oscillations or quiescent states, have been observed in various nervous systems, but the dynamics of this event remain unclear. Here, we report that on-off firing patterns observed in three experimental models (rat sciatic nerve subject to chronic constrictive injury, rat CA1 pyramidal neuron, and rabbit blood pressure baroreceptor) appeared as an alternation between quiescent state and burst containing multiple period-1 spikes over time. Burst and quiescent state had various durations. The interspike interval (ISI) series of on-off firing pattern was suggested as stochastic using nonlinear prediction and autocorrelation function. The resting state was changed to a period-1 firing pattern via on-off firing pattern as the potassium concentration, static pressure, or depolarization current was changed. During the changing process, the burst duration of on-off firing pattern increased and the duration of the quiescent state decreased. Bistability of a limit cycle corresponding to period-1 firing and a focus corresponding to resting state was simulated near a sub-critical Hopf bifurcation point in the deterministic Morris-Lecar (ML) model. In the stochastic ML model, noise-induced transitions between the coexisting regimes formed an on-off firing pattern, which closely matched that observed in the experiment. In addition, noise-induced exponential change in the escape rate from the focus, and noise-induced coherence resonance were identified. The distinctions between the on-off firing pattern and stochastic firing patterns generated near three other types of bifurcations of equilibrium points, as well as other viewpoints on the dynamics of on-off firing pattern, are discussed. The results not only identify the on-off firing pattern as noise-induced stochastic firing pattern near a sub-critical Hopf bifurcation point, but also offer practical indicators to discriminate bifurcation types and neural excitability types.
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Action Potentials , Pyramidal Cells/physiology , Stochastic Processes , Animals , Male , Models, Biological , Rabbits , Rats , Rats, Sprague-DawleyABSTRACT
Objective To study the effects of bFGF on the proliferation of cultured human melanocytes, and to seek a quick method for in vitro culture of human melanocytes. Methods Melanocytes were isolated from human foreskin, and divided into two parts to be cultured with or without the presence of bFGF (0.3 μg/L). Second-passage melanocytes were identified with immunochemical stain. The growth of melanocytes was observed every 3 days for 12 days. Third-passage melanocytes were treated with various concentrations (0.3 - 2.1 μg/L) of bFGF for 72 hours followed by the detection of proliferation of and trosinase activity in melanocytes. Results Human melanocytes were obtained from primary culture in medium containing certain concentrations of bFGF, which were identified with immunohistochemical stain. The morphology of cultured melanocytes varied with growth stage of cells. The bFGF-treated melanocytes appeared to grow more rapidly than untreated melanocytes. Further more, a significant increase was observed in the proliferation rate of melanocytes treated with bFGF of 0.3 and 0.6 μg/L (P<0.05 or 0.01 ) and tyrosinase activity in melanocytes treated with bFGF of 1.5 and 1.8 μg/L (P < 0.05 or 0.01 ) in comparison with the untreated melanocytes.Conclusions The addition of certain concentrations of bFGF to defined medium can benefit the primary culture of melanocytes and make it possible to get large quantities of purified melanocytes with high viability in short periods. Certain concentrations of bFGF can up-regulate the proliferation of and tyrosinase activity in melanocytes.
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@#ObjectiveTo observe the protective effect of high-dosage methylprednisolone (MP) after lumbar spinal stenosis operation. Methods20 Patients with lumbar spinal stenosis accepted laminectomy. The treatment group (11 cases) received MP 30 mg/kg when the skin incised, and followed with MP 40 mg twice a day for 3 d after operation. The control group (9 cases) received MP 40 mg twice a day for 3 d after operation. They were assessed with JOA scale before and after operation. ResultsThe JOA score was (9.25±2.12) and (9.53±2.10) in treatment group and control group before operation (P>0.05), and was (13.43±2.01) and (11.21±2.13) 1 week after operation (P<0.05), (14.62±2.15) and (13.04±2.11) 3 months after operation (P<0.05). The neurological symptom deteriorated in 1 cases of control group. No peptic ulcer, poor wound healing or other associated complications has been found. ConclusionHigh-dosage methylprednisolone may protect the spine cord intraoperative the lumbar spinal stenosis.
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Objective To investigate the relationship of fascin-1 protein expression with the metastasis of basal cell carcinoma and squamous cell carcinoma. Methods Skin specimens were obtained from 10 normal human controls, 13 patients with basal cell carcinoma (8 nodular variant and 5 superficial variant) and 24 patients with SCC (11 SCC in situ and 13 invasive SCC). Immunohistochemical staining was performed to analyze the expression of fascin-1 protein. The staining results were quantitatively assessed with computer image analysis system (Image-pro Plus 6.0). Results The optical density of fascin-1 averaged 0.1152±0.04574 in SCC in situ, 0.1257±0.03096 in invasive SCC, and 0.0293±0.00981 in normal controls; the expression of fascin-1 was significantly higher in SCC tissue than in normal control skin (both P<0.05). Increased optical density was also observed for fascin-1 in nodular variant of SCC (0.0808 ±0.05642) and superficial variant of SCC (0.0806±0.04346) compared with the normal controls, whereas no statistical difference was observed between nodular and superficial variant of SCC. Conclusion In BCC and SCC, there is an over expression of fascin-1, which may be linked to the local invasion of carcinoma,
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Objective To study the in vitro culture condition for melanoblasts from human foreskin tissue. Methods The skin tissue taken from foreskin of children was treated with 0.5% dispase Ⅱ to separate epidermis from dermis, then with trypsin to obtain single cell suspension, which was cultured in modified medium for melanoblasts, i.e., MCDB254 medium supplied with several cell growth factors. Finally, melanoblasts were obtained based on the difference of adhesion speed. The morphology and proliferation of cultured melanoblasts were observed under a light microscope. DOPA staining, immunostaining with anti- S-100 and -tyrosinase related protein 2 (TRP2) antibodies, and transmission electron microscopy were per- formed to identify the cultured melanoblasts. Results The cultured human melanocytes displayed a match-like shape, scattered arrangement, syrmnetric double poles, slim cell body, highly refractive nuclei; meanwhile, the melanoblasts exhibited plentiful cytoplasm, large volume, bipolar or irregular shape and clonal growth. Additionally, the melanocytes were positive for TRP2, S-100 and Dopa staining, while the melanoblasts were positive only for TRP2. Electron microscopy revealed the presence of mature melanin granules (stage Ⅲ-Ⅳ ) in melanocytes but immature melanin granules (stage Ⅰ ) in melanoblasts. Conclu- sion Stable pure culture of melanoblasts has been realized with the reformed medium, which may lay a foundation for the investigation into the mechanism of epidermal pigmentation.
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Objective To construct tissue-engineered skin via in vitro inoculation of epidermal stem cells(ESCS)onto de-epidermized dermis.Methods Skin tissue was obtained from the foreskin of a healthy 6-year-old child.and keratinocytes were isolated by two-step trypsinization method followed by the collection of ESCS via rapid adhesion by collagen Ⅳ.The ESCS were identified by morphological observation and immunohistochemical staining with K19 and integrin β1.To construct tissue-engineered skin,selected ESCS were seeded onto the surface of de-epidermized dermis followed by a one-week culture immersed in the medium and a subsequent 4-week culture at the air-medium interface.The tissue-engqneered skin was evaluated with haematoxylin & eosin(HE)staining as well as keratin immunohistochemistry.Results Micro scopically,cultured ESCs showed a paving stone-like appearance and grew into colonies.Immunohistochemistry revealed that the ESCs were positive for integrin-β1 and keratin 19.After 5 weeks of culture,3-6 layers of epidermal cell were observed on the dermis with the formation of stratum corneum.Keratin protein was observed in the artificial epidermal skin.Conclusion Tissue-engineered skin is successfully constructed with epidermal stem cells and de-epidermized dermis in vitro.
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Objective To investigate the phenotype,number and distribution of inflammatory cells in early and late stages of spontaneous regression of halo nevi,and to elucidate the immunological mechanisms for spontaneous regression of these nevi.Methods Halo nevi,their surrounding non-lesional skin,and normal control skin were examined by immunohistochemical staining with monoclonal antibodies to CD3,CD4,CD8,CD20,CD1a,CD56 and CD68.Staining results were observed and analyzed by the computer image analysis system,image-pro plus 6.0.Results The number of CD4+,CD8+,CD20+,CD1a+cells,along with the diameter of CD1a+and CD68+ cells was significantly increased in the lesions of early and late stage of spontaneous regression of halo nevi than in non-lesional skin and normal control skin(both P<0.01).The ratio of CD8+/CD4+ cells in the lesions of late stage of spontaneous regression was also higher than that in the lesions of early stage (2.05∶1 VS 1.82∶1).A massive infiltrate of CD8+ cells was observed in the nests of nevus cells.ConclusionsCD4,CD8,CD20,CD1a,CD56 and CD68 positive cells are all involved in the spontaneous regression of halo nevi,and CD8+ cells may play a predominant role in this process.
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Objective: To study the influence of Centipede Acidic Protein(CAP) on the proliferation and collagen synthesis of cultured neonatal rat cardiac fibroblasts(CFb) induced by angiotensinⅡ(AngⅡ),and to explore the mechanisms of CAP on cardiac fibrosis.Methods: Neonatal rat cardiac fibroblasts were treated with AngⅡ to produce fibrosis model.The effects of CAP on proliferation of CFb were observed by MTT colorimetric assay,synthesis of collagen was observed by the hydroxyproline concentration.The NO contents were measured by Nitric acid reductase method.The c-myc expression was examined by semi-quantitative RT-PCR analysis.Results: Compared with that of control group,the proliferation,collagen synthesis and the levels of c-myc mRNA expression of CFb in the model group increased,while the NO contents decreased obviously(P