OBJECTIVETo construct and characterize the TGF-β1, induced epithelial-mesenchymal transition (EMT) model of keloid epithelial cells in vitro, and to investigate the expression of epithelial stem cells related surface markers in keloid epithelial cells during EMT induction.

METHODSThe epithelial cells from 3 keloid samples of ears were cultured in vitro and induced by transforming growth factor betal (TGF-β1, 1 ng/ml) for 5 days, which was the experimental group, the same cells untreated were considered as the negative control group. The expressions of EMT-associated markers and regulative genes were detected using immunofluorescence staining, real-time PCR and western blot analysis. Then the surface markers of epithelial stem cells were detected using real-time PCR. Statistical significance was determined using Independent-Samples t Test, a p value less than 0. 05 was considered statistically significant.

RESULTSThe mRNA expression of transcription factor snail2 and mesenchymal-specific marker vimentin increased significantly in TGF-β1, induced keloid epithelial cells (P < 0. 05), in which snail2 increasing from 0. 91 ± 0. 23 to 1. 69 ± 0. 10, and vimentin from 5. 86 ± 2. 07 to 24. 29 ± 5. 39. Whereas the mRNA expression of epithelial-specific marker E-cadherin decreased from 1. 06 ± 0. 19 to 0. 65 ± 0. 09. The mRNA expression of CD29 and Lgr6, two surface markers of epithelial stem cells, significantly increased after induction of the TGF-β1, (P < 0. 05), from 0. 55 ± 0. 14 and 1. 61 ± 0. 31 to 1. 19 ± 0. 12 and 3. 84 t 0. 62 respectively. In induced cells, the immunofluorescence results showed staining of E- cadherin became faint, but the number of positive staining cells of vimentin increased. Western blot confirmed the protein expression of E-cadherin weakened, and the vimentin and p-Smad3 enhanced (P < 0. 05).

CONCLUSIONSTGF-β1, initiated EMT in keloid epithelial cells by inducing the up-regulation of snail2, and TGF-β1,/Smad3 signaling pathway was involved in EMT. EMT could change the phenotype of epithelial stem cells in keloid.

Biomarkers ; metabolism ; Cadherins ; genetics ; metabolism ; Epithelial Cells ; drug effects ; physiology ; Epithelial-Mesenchymal Transition ; drug effects ; physiology ; Humans ; In Vitro Techniques ; Keloid ; pathology ; RNA, Messenger ; metabolism ; Signal Transduction ; Smad3 Protein ; genetics ; metabolism ; Snail Family Transcription Factors ; Transcription Factors ; genetics ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; pharmacology ; Up-Regulation ; Vimentin ; genetics ; metabolism

Objective: To observe the time-effect relationship of moxibustion for primary dysmenorrhea (PD) due to stagnation and congelation of cold-damp, thus explore the optimal choice of moxibustion duration, and provide evidence for achieving satisfactory efficacy in moxibustion treatment. Methods: A total of 90 patients with PD due to stagnatin and congelation of cold-damp were divided into three groups by the random number table method, with 30 cases in each group. All the patients in the three groups were given moxibustion treatment at Guanyuan (CV 4), 20 min in group A, 40 min in group B and 60 min in group C. The changes in the pain measurement score in the three groups were observed after treatment. Results: After treatment, there were significant differences in the clinical efficacy among the three groups (P<0.05); the clinical efficacy was better in group B and group C than that in group A (P<0.05), and that in group B was better than that in group C (P<0.05). Besides, the pain measurement score changed significantly after treatment in the three groups (all P<0.05), and the between-group differences were also statistically significant (P<0.05); the pain measurement scores in group B and group C were lower than that in group A (P<0.05), and that in group B was lower than that in group C (P<0.05). Conclusion: Given the same stimulating frequency and intervention time of moxibustion, 40-minute duration demonstrates relatively better efficacy for PD due to stagnation and congelation of cold-damp.

OBJECTIVETo study the relations between anthropometric parameters [body mass index (BMI), percent of body fat, waist circumference] and blood lipids of urban children in Beijing and to compare the ability of prediction effect of waist circumference, percent body fat and BMI on lipid profiles.

METHODS852 children aged 9-10 were recruited with multi-stage sampling. Serum lipids of children were measured using automatic biochemical instrument. Fisher's exact test, trend test and median regression were used to analyze the data from 847 subjects with valid information.

RESULTS(1)With the rise of the BMI, percent body fat and waist circumference, total triglycerides (TG), total cholesterol (TC) and low density lipoprotein choleasterol (LDL-C) showed increasing trends while high density lipoprotein choleasterol (HDL-C) had a decreasing trend(P < 0.01). (2)TG was mainly influenced by waist circumference (t = 6.86), sex (t = 3.96) and percent of body fat (t = 1.85); TC was influenced by percent of body fat (t = 5.31); LDL-C was influenced by waist circumference (t = 4.65), height (t = -2.79) and percent age of body fat (t = 2.77); HDL-C was influenced by waist circumference (t = -12.24), sex (1= -5.83).

CONCLUSIONWith the aggravation of children's fatness (BMI, WC and PBF), serum lipids would increase and WC seemed to be the best predictor for lipids among BMI, WC and PBF.

Body Composition ; Body Height ; Body Mass Index ; Child ; China ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Humans ; Triglycerides ; blood ; Urban Population ; Waist Circumference

OBJECTIVETo explore the role of stem cell mobilization on regeneration of partially grafted livers.

METHODSRats models with cross-sex 50% PLTx (partial liver transplantation) were established. The rats were divided into three groups: PLTx, WLTx (whole liver transplantation) and sham operation groups. Bone marrow and liver samples were collected on days 1, 3, 5, 7 postoperatively (each n = 6). The quantitative variations of the cells with stem cell markers in the bone marrow, including beta2m-/Thy-1.1+, CD45+/CD34+, Flt2/3+ and c-kit+ markers, were detected using flow cytometry. Sry gene positive cells in donor livers were detected by fluorescent in situ hybridization (FISH), and the expressions of CD34, c-kit and Thy-1.1 were detected by immunohistochemistry technique.

RESULTSCompared with the WLTx and sham operation groups, beta2m-/Thy-1.1+, CD45+/CD34+ cells in bone marrows in the PLTx group increased on the first postoperative day and decreased on the following days. The CD34, c-kit and Thy-1.1 positive cells detected in portal tract areas peaked during the 3-5 postoperative days. CD34+/CD45+ positive cells could be detected. The expressions of CD34, c-kit and Thy-1.1 positive cells were rare in the WLTx and sham operation groups. Sry+ cells could be detected in portal tract areas and few Sry+/CD34+ and Sry+/Thy-1.1+cells were detected.

CONCLUSIONIn the PLTx group, the stem cells in the bone marrow were mobilized and stem cells in the liver were activated.

Animals ; Antigens, CD34 ; immunology ; Bone Marrow Cells ; cytology ; immunology ; Female ; Hematopoietic Stem Cell Mobilization ; Leukocyte Common Antigens ; immunology ; Liver Transplantation ; methods ; Male ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; immunology ; Thy-1 Antigens ; immunology

Objective: To treat the depressed scars by injecting nanofat and investigate its therapeutic effect. Methods: Autologous fat was harvested from abdomen or thigh using low-pressure suction. The lipoaspirate was mechanically emulsified after rinsing. Emulsification of the fat was achieved by shifting the fat between two 5 ml syringes connected to each other by a three direct connector. After this emulsification process, the fatty liquid was again filtered over the sterile nylon cloth. Nanofat was injected into the dermis of depressed scars using a 26-gauge needle and the injection volume was 1-2 ml/cm². After three months, another injection would be performed if the depressed scar remained obvious. Results: From January 2016 to October 2017, eighteen patients and thirty-three depressed scars were treated. There was a temporary erythema of the injected area that lasted two to three weeks. The clinical result gradually improved over time and were maximal from three months postoperatively for most cases. Three months after nanofat injecting, the cavity of scars was significantly decreased; The color of scars were significantly improved and more close to the adjacent skin; The stiffness of scars was also obvious decreased. The follow-up ranged 4 months to 18 months and the average was 11.0±4.6 months. Seventeen patients were satisfied with the result, one patients was not satisfied and the satisfaction rate was 94%. No infections, fat cysts, granulomas, or other unwanted side effects were observed. Conclusions Nanofat injecting is a definite and effective treatment for depressed scars with fewer complications.

Objective: To clarify the status of proliferation and differentiation of RCJ3.1C5.18 cells in chondrogenesis and explore insulin-like growth factor binding proteins (Igfbps) gene expression profile during the process. The results provided a research foundation for understanding the role of Igfbps in chondrogenesis. Methods: Fetal rat derived mesenchymal chondrogenic cell line RCJ3.1C5.18 was used, which progresses spontaneously to differentiated growth plate chondrocytes. The proliferation and differentiation status of cells were determined by cell staining, real-time PCR and Muse™ Cell Analyzer at days 1, 4, 7 and 10 of culture respectively. Igfbps gene expression was assessed by real-time PCR. Results: In the early period (1-4 days), cells proliferated rapidly and the expression of chondrocytes marker genes, such as typeⅡ collagen, SRY-box 9 and aggrecan increased gradually. Igfbp4 and Igfbp6 gene expression paralleled the expression of chondrocytes marker genes. On days 7-10 of culture, cells viability decreased and gradually developed hypertrophic-like and osteogenic-like characteristics, which was confirmed by specific staining and gene expression of alkaline phosphatase, matrix metallopeptidase 13.Igfbp1, Igfbp3 and Igfbp5 mRNA levels were up regulated gradually at this stage. Conclusions The status of proliferation and differentiation of cells are different in the process of chondrogenesis. Changes in Igfbp gene expression are associated with chondrocyte differentiation, suggesting that they have different role during chondrocyte proliferation and differentiation.

Background: Increased right ventricle afterload during acute respiratory distress syndrome(ARDS)may induce acute cor pulmonale(ACP),which is associated with a poor clinical outcome.Echocardiography is now considered as a rapid and non-invasive tool for diagnosis of ACP.The aims of this study were to investigate the morbidity and mortality rates of ACP in ARDS patients in intensive care units(ICUs)across the mainland of China and to determine the severity and prognosis of ACP in ARDS patients through an ultrasound protocol(TRIP).And the association between ACP related factors and the ICU mortality will be revealed.Methods: This study is a multicenter and cross-sectional study in China which will include ICU participants when diagnosed as ARDS.The ultrasound protocol,known as the TRIP,is proposed as severity assessment for ACP,which includes tricuspid regurgitation velocity(T),right ventricular size(R),inferior vena cava diameter fluctuation(I),and pulmonary regurgitation velocity(P).The 28-day mortality,ICU/hospital mortality,the length of stay in ICU,mechanical ventilation days,hemodynamic parameters and lab parameters of liver function and kidney function are all recorded.Discussion: This large-scale study would give a sufficient epidemic investigation of ACP in ARDS patients in China.In addition,with the TRIP protocol,we expect that we could stratify ACP with more echocardiography parameters.

OBJECTIVETo investigate the efficacy of continuous propofol infusion via the common carotid artery for general anesthesia.

METHODSForty adult patients scheduled for abdominal surgery were randomly assigned into 2 groups to receive propopol via the common carotid artery (IC group, n=20) or via the median cubital vein (IV group, n=20). Anesthesia was induced with intravenous administration of drugs and maintained with continuous propofol infusion via the common carotid artery or the median cubital vein, with the CSI stabilized at 40-/+5 till the end of the operation. During the anesthesia, intravenous injection of fentanyl (3 microg.kg(-1).h(-1)) and vecuronium (50 microg.kg(-1).h(-1)) were given intermittently to maintain the analgesia and muscular relaxation. The dose of propofol used, hemodynamics and recovery of the patients were observed.

RESULTSThe dose of propofol used during the surgery to maintain a CSI of 40-/+5 was significantly lower in group IC and than in group IV (2.57-/+0.67 vs 5.72-/+1.37 mg.kg(-1).h(-1), P<0.01). In group IC, the blood pressure was elevated in more than half of the patients and in some cases, the elevation exceeded one third of baseline value and needed intervention with hypotensive drugs. In the IV group, the patients' blood pressure remained stable and varied within the amplitude of 15% of the baseline level. Recovery of spontaneous breathing and consciousness was more quickly in group IC than in group IV (P<0.05).

CONCLUSIONLoss of consciousness and nervous reflex can be achieved with propofol infusion via the common carotid artery, which reduces propofol dose by about 50% in comparison with intravenous infusion and allows more rapid recovery of spontaneous breath and consciousness.

Abdomen ; surgery ; Adult ; Aged ; Analgesics, Opioid ; administration & dosage ; Anesthesia, General ; methods ; Carotid Artery, Common ; Female ; Fentanyl ; administration & dosage ; Humans ; Hypnotics and Sedatives ; administration & dosage ; Infusions, Intra-Arterial ; Male ; Middle Aged ; Nicotinic Antagonists ; administration & dosage ; Propofol ; administration & dosage ; Treatment Outcome ; Vecuronium Bromide ; administration & dosage

OBJECTIVETo investigate the cytotoxic effect of epigallocatechin gallate (EGCG) on human hepatocellular carcinoma cell line HepG2 cells and corresponding changes of TGF-beta1-Smad pathway.

METHODSThe cytotoxic effect of EGCG on HepG2 cells was determined by MTT assay. Cell cycle and apoptosis rate were detected by flow cytometry. RT-PCR and luciferase assay were used to verify whether TGF-beta1-Smad signaling pathway is intact in HepG2. The mRNA expression of Smad 2, Smad3, Smad4 and Smad7 was detected by real-time PCR.

RESULTSEGCG induced apoptosis in the HepG2 cells in a time- and concentration-dependent manner. The proportion of G(1) phase cells was increased gradually as the concentration increased. However, the percentage of cells in S phase was decreased gradually. Annexin V/PI assay demonstrated that early apoptosis increased as the concentration increased, and late apoptosis also increased, when treated with high-concentration EGCG. The intact TGF-beta1-Smad pathway was verified by luciferase assay and RT-PCR. There was no significant effect of EGCG on mRNA level of Smad 2, Smad 3, and Smad 4 in HepG2 cells, but downregulated mRNA level of Smad 7.

CONCLUSIONEGCG can reduce apoptosis in human hepatocellular carcinoma cell line HepG2 cells. The activation of TGF-beta1-Smad signaling pathway may be involved in its cytotoxicity mechanisms.

Anticarcinogenic Agents ; pharmacology ; Apoptosis ; drug effects ; Catechin ; analogs & derivatives ; pharmacology ; Cell Cycle ; drug effects ; Hep G2 Cells ; Humans ; RNA, Messenger ; metabolism ; Signal Transduction ; Smad Proteins ; genetics ; metabolism ; Smad7 Protein ; genetics ; metabolism ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo investigate the effect of apelin on vasodilatation of isolated pulmonary arterial rings in rats and its relationship to the nitric oxide (NO) pathway, and to observe the difference of vasodilatation between hypoxic rats and normoxic rats.

METHODSThirty-six male Sprague-Dawley (SD) rats were randomly divided into hypoxic group and normoxic group. The effects of accumulated apelin on pulmonary arterial rings preconstricted with norepinephrine (NE) were observed by using tissue organ bath system. After pulmonary arterial rings were pretreated with three methods: removing the endothelium, pretreating with nitric oxide synthase inhibitor L-NAME or soluble guanylatecyclase inhibitor ODQ, the different effect of apelin was observed. In addition, the difference of vasodilatation between hypoxic rats and normal rats were observed.

RESULTS(1) Exposure of intact endothelium pulmonary arterial rings preconstricted by NE to apelin at concentration (0.01 - 100 nmol/L) induced a significant concentration dependent relaxation. The maximal vasorelaxant effect of apelin was 10.62% +/- 2.60%, which was inhibited by removal of the endothelium (P < 0.01), pretreatment with L-NAME (P < 0.01) or ODQ (P < 0.01). (2) Response of pulmonary arterial rings from hypoxic pulmonary hypertension rats was decreased (P < 0.05). Compared to normal rats, at a concentration of 100 nmol/L, the response to apelin on arteries from hypoxic rats decreased 60.45% (P < 0.01). But the values of EC50 were not significantly different (P > 0.05).

CONCLUSIONThese results indicate that apelin relaxes the pulmonary arterial rings of rats in an endothelium dependent manner, which may have a relationship to NO signaling pathway. The response of vasodilatation is decreased in the pulmonary arterial rings from the hypoxic rats.

Animals ; Apelin ; Hypoxia ; physiopathology ; In Vitro Techniques ; Intercellular Signaling Peptides and Proteins ; pharmacology ; Male ; Nitric Oxide ; metabolism ; Pulmonary Artery ; physiopathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Vasodilation ; drug effects