BACKGROUND: Transforming growth factor beta (TGF-β) has an important role in tendon healing and adhesion formation.Inhibiting TGF-β and its receptor expression may prevent adhesions after tendon open.OBJECTIVE: To study the effects of mannose-6-phosphate, a natural inhibitor of TGF-β, on TGF-β and its receptor production in tendon sheath fibroblasts, epitenon tenocytes, and endotenon tenocytes of rabbit flexor toes.METHODS: Tendon sheath fibroblasts, epitenon tenocytes, and endotenon tenocytes were isolated from rabbit flexor tendon and cultured separately. All these cells were divided into 2 groups at random, experiment group supplemented with mannose-6-phosphate and control group without mannose-6-phosphate. The expression of TGF-β and TGF-β receptor was quantified with enzyme-linked immunosorbent assay. The expression of TGF-β1 was also assessed with in situ hybridization and immunohistochemistry.RESULTS AND CONCLUSION: The expression of TGF-β and TGF-β receptor in experiment group was significantly lower than that in control group (P ＜ 0.05). In experimental group, the positive expression of TGF-β1 mRNA and the expression level of intracellular TGF-β1 mRNA in all tendon cells demonstrated significantly lower than those in the control group (P ＜ 0.05).Immunohistochemical staining showed expression of TGF-β1 were significantly lower in all three types of tendon cell cultured with mannose-6-phosphate.

BACKGROUND:Mesenchymal stem cel s are the seed cel s for tendon tissue engineering which can be obtained in large quantities, but how to induce in vitro is a key technology.
OBJECTIVE:To explore the effect of platelet-rich plasma on the proliferation and col agen production of in vitro cultured mesenchymal stem cel s.
METHODS:The rabbit mesenchymal stem cel s were separated ad cultured. The high-dose platelet-rich plasma group, middle-dose platelet-rich plasma group and low-dose platelet-rich plasma group were set to induce the mesenchymal stem cel s, and the blank control group was set as control.
RESULTS AND CONCLUSION:The proliferation of mesenchymal stem cel s in the high-dose platelet-rich plasma group, middle-dose platelet-rich plasma group and low-dose platelet-rich plasma group was high, and in rapid growth with big increase amplitude, and there was no significant difference in the proliferation when compared with the blank control group (P<0.05). The effect was positively correlated with the culture time, and after cultured for a certain time, the effect was in dose-dependent manner, as higher dose platelet-rich plasma had more significant effect on the proliferation of the cel s. The results indicate that platelet-rich plasma can significantly promote the synthesis of col agen type Ⅰ and Ⅲ of mesenchymal stem cel s, the higher the dose, the more significant the effect on the col agen.

AIM: To study the anti-invasive effect of NS-398 on colon cancer cell line HT-29 in vitro an its regulation by CD44v6 and nm23-H1 genes. METHODS: Flow cytometry was used to detect the expression of COX-2 and CD44v6 in HT-29 cells. MTT was used for cell survival rate tests. The modified Boyden chamber model was used for quantitative invasion assay. RT-PCR was used to detect the expression of nm23-H1 mRNA. RESULTS: Flow cytometry analysis showed that COX-2 was positive in HT-29 cells. NS-398 had significant inhibitory effects on invasion of HT-29 cells, which had no relation with its cytotoxicity. NS-398 down-regulated the expression of CD44v6 and up-regulated the expression of nm23-H1 mRNA. CONCLUSION: NS-398 has an anti-invasive effect on HT-29 cells in vitro. Down-regulation of CD44v6 and up-regulation of nm23-H1 may be its underlying mechanisms.

Objective To detect anti-clinically amyopathic dermatomyositis (CADM)-140 antibody in patients with dermatomyositis (DM) or CADM,and to estimate its clinical correlation.Methods Serum samples were collected from 22 patients with DM,16 patients with CADM,46 patients with other connective tissue diseases complicated by interstitial lung disease(including 8 cases of polymyositis,15 cases of systemic lupus erythematosus,5 cases of systemic sclerosis,6 cases of Sj(o)gren syndrome,6 cases of mixed connective tissue disease,6 cases of idiopathic pulmonary fibrosis),and 5 normal human controls.Enzyme-linked immunosorbent assay (ELISA) was performed with the recombinant melanoma differentiation-associated gene 5(rMDA)as a substrate to measure the anti-CADM-140 antibody in these serum samples.Clinical manifestations were compared between patients with anti-CADM-140 antibody and those without.Results The anti-CADM-140antibody was found in 43.8% (7/16) of patients with CADM and 9.1%(2/22) of patients with DM(P<0.05),but absent in the patients with other connective tissue diseases and in the normal human controls.A significant incroase was observed in anti-CADM-140 antibody-positive patients with DM/CADM in the incidence of cutaneous ulceration and necrosis,interstitial lung disease and rapidly progressive interstitial lung disease (8/9 vs.6.9%,P<0.01;9/9 vs.48.3%,P<0.01;5/9 vs.0,P<0.05),serum lactate dehydrogenase level(328.3±104.2 vs 241.1±100.3 IU/L P<0.05),erythrocyte sedimentation rate(40.8±23.1 vs.22.5±16.8 mm/1 h,P<0.05),high resolution computed tomography score(122.9±54.8 vs.70.0±59.8,P<0.05)compared with anti-CADM-140 antibody-negative patients with DM/CADM.The ereatine kinase level was significantly lower(156.3±260.8 vs.1806.2±3737.1 IU/L P<0.05)in anti-CADM-140 antibody-positive patients with DM/CADM than in anti-CADM-140 antibody-negative patients with DM/CADM,while no significant difference was noted in the positivity rate of antinuclear antibodies or incidence of malignancies between the antibody-positive and-negative patients with DM/CADM.Conclusions Anti-CADM.140 antibody not only is useful for the diagnosis of interstitial lung disease in patients with DM/CADM,but also may serve as a serum marker for rapidly progressive interstitial lung disease.Monitoring of serum anti-CADM-140 antibody might help to predict the progression of interstitial lung disease in patients with DM/CADM.

Objective To observe the effects of genistein on PCNA expression and cell cycle in fibroblasts derived from human hypertrophic scar in order to explore the mechanism of its inhibition on hypertrophic scar (HS) fibroblast proliferation. Methods The human hypertrophic scar fibroblasts were cultured in vitro. Genistein with various concentrations (25, 50, 100 μmol/L) was co-cultured in the medium for 48 hours. The expression of PCNA was detected with immunocytochemical staining method and the cell cycle was measured with flow cytometry. Results Genistein could significantly decrease PCNA expression in HS fibroblasts, especially when its concentration at 50 μmol/L or 100 μmol/L. The cell percentage of G0～G1 phase decreased with drug′s concentration, and G2～M percentage increased conversely, implying the suspension of mitosis. In 100 μmol/L group, most cells blocked at S phase and a hypodiploid apoptosis peak could be observed ahead of G1 phase. Conclusion Genistein can inhibit the proliferation of human hypertrophic scar by blocking cell division as well as decreasing DNA synthesis.

Objective To evaluate liver biopsy for the diagnosis in liver transplant patients with suspected acute rejection. Methods From Oct. 2004 to Apr. 2005, liver biopsies were performed 53 times in 39 transplant cases. Results Based on Banff schema for grading liver allograft rejection, laboratory abnormalities and result of treatment, acute rejection was diagnosed on 16 episodes, preservation injury in 12, bile duct strictures in 9, drug-induced injury in 11, chronic rejection in 3 and acute hepatic failure in 2. Conclusions Hepatocyte ballooning with necrosis features preservation injury. Drug-induced injury commonly has a combination of hepatocyte denaturalization with mild portal inflammation. Histologic features of early bile duct strictures in liver biopsy show prominent bile ductular proliferation and the canalicular cholestasis with mild hepatocyte damage which help to exclude acute rejection.

Objective To summarize the key points of nursing care in reservation of catheters in 11 cases of nasopharyngeal carcinoma patients who received concurrent chemo-radiotherapy with symptomatic thrombosis after ultrasound-guided implantation of double-lumen PICCs(PowerPICC).Methods From January,2014 to December,2015,totally 11 cases with symptomatic venous thromboembolism were identified among 109 cases of nasopharyngeal carcinoma patients receiving concurrent radiotherapy and chemotherapy,and observation and nursing care were provided at the early stage of thrombosis and during thrombosis.Results All double-lumen PICCs were reserved,and no recurrence or aggravation of thrombosis was recorded to the end of the treatment.The duration of carrying PICCs was 67～89(77.45±6.65) days.Conclusion With careful treatment and nursing,PICC catheter-related complications can be reduced and the duration of carrying catheters can be prolonged,which leads to accomplishment of the treatment plan for patients.

Objective To investigate the estimated values of sequential organ failure assessment (SOFA) and quick SOFA (qSOFA) for diagnosis and prognosis in patients with sepsis according to the new diagnostic criteria in Sepsis 3.0.Methods A retrospective study was conducted.All the clinical data were collected from patients with definite diagnosis of infection and they were admitted into the Intensive Care Unit (ICU) of Beijing Traditional Chinese Medicine Hospital Affiliated to Capital Medical University from July 2014 to June 2016.The patients' gender,age,infectious location,respiratory rate (RR),oxygenation index (PaO2/FiO2),Glasgow coma scale (GCS),total bilirubin (TBil),platelet count (PLT),serum creatinine (SCr),serum lactate level,etc.general data on admission were collected to carry out SOFA and qSOFA scorings.And then the septic patients in accord with the diagnostic criteria of Sepsis 3.0 were screened out.According to outcome after admission,the septic patients were divided into survival group and death group,and the differences in diagnosis and in estimation value of prognosis between SOFA scoring and qSOFA scoring were assessed as SOFA group and qSOFA group.Results From 545 septic patients enrolled,189 septic patients consistent with the diagnostic criteria of Sepsis 3.0 were selected.In SOFA scoring group,the morbidity of septic patients was 34.68％,while in qSOFA scoring group,it was 15.96％,the difference between the two groups being statistically significant (P ＜0.01).The mortality was significantly lower in SOFA scoring group than that in qSOFA scoring group [28.04％ (53/189)vs.42.53％ (38/87),P ＜ 0.05].The mortality of qSOFA scoring group was about 1.52 times that of SOFA scoring group.On the aspect of scoring,in patients with SOFA scoring the score of death group was significantly higher than that in survival group (8.74 ± 0.417 vs.7.10 ± 0.235,P ＜ 0.01);in the patients with qSOFA scoring,the score in death group compared with that in survival group showed uo statistical significant difference (2.32 ± 0.48 vs.2.16 ± 0.37,P ＞ 0.05).On the aspect of laboratory indexes,the levels of GCS score in death group was significantly lower than that in the survival group (8.15 ± 0.67 vs.12.48 ± 0.36),blood lactate level in death group was significantly higher than that in the survival group (mmol/L:8.55 ± 4.66 vs.2.31 ± 0.16,P ＜ 0.01);the PaO2/FiO2,TBil,PLT and SCr showed no significant differences between the two groups (all P ＞ 0.05).Conclusions The new diagnostic criteria (Sepsis 3.0) can be used for diagnosis of sepsis in ICU.Compared with qSOFA scoring,the SOFA scoring is more suitable to be used for diagnosis and predicting prognosis of septic patients in ICU;SOFA scoring,GCS scoring and serum lactate level can be applied to estimate outcome of septic patients.

AIM: To observe the change of subunit of NADPH oxidation enzyme complex nox - 1 protein in cardiocyte hypoxia - reoxygenation injury and the role of cardiotrophin -1.METHODS: Cardiomyocytes from the hearts of 1 -3 d old neonatal rats were prepared by a modified method. Five groups were included in the study: control; hypoxia/ reoxygenation; hypoxia/reoxygenation + CT - 1; CT - 1 + hypoxia/reoxygenation + LY294002 (PIK3/Akt inhibitor) ; CT -1 + hypoxia/reoxygenation + PD98059 (ERK inhibitor) ; CT - 1 + hypoxia/reoxygenation + DMSO. The concentration of CT -1 was 10 μg/L. The survival rate of myocytes was evaluated by MTS method. Apoptosis, mitochondrial permeability transition pore ( △ψm) and reactive oxygen species ( ROS) were detected by flow cytometry. Nox - 1 protein was determined by Western blotting. RESULTS: Apoptosis of cardiomyocytes and the level of ROS (19.7% ±1.4% vs 2.1% ± 0.5% , 14.07% ± 1.25% vs 3.54% ± 0.86% , P < 0.05 ) increased markedly after hypoxia/reoxygenation, but cardio-myocyte survival rate and the level of△ψm (40.55% ±4.25% vs 86.28% ±7.15% , P <0.01) decreased significantly. The expression of nox - 1 protein was upregulated markedly. With CT - 1 intervention, cardiomyocyte survival rate increased markedly, apoptosis, both ROS and expression of nox - 1 protein reduced significantly. The level of△ψm increased obviously. The effect of CT - 1 was inhibited by LY294002.No significant effect was observed on cells survival in DMSO group, which confirmed that LY294002 was specifically involved in blocking the protective effect of CT - 1.CONCLUSION : The expression of subunit of NADPH oxidation enzyme complex nox - 1 protein is upregulated markedly in cardiocyte hypoxia - reoxygenation injury.CT - 1 protects cardiac cells against hypoxia - reoxygenation injury by downregulating the expression of nox -1 protein to decrease the level of ROS.

Objective To study the effect of Cardiotrophin-1 (CT-1) on cardiocyte hypoxia-reoxygenation injury,and to investigate the signaling pathways involved in the protective effect. Method This study was carried out in Key Lab of Molecular Medicine in Jiangxi Province. Cardiomyocytes from the hearts of 2-day-old Sprague-Dawley neonatal rats were prepared by a modified method. Five groups were included in the study. Group (ⅰ): control, Group (ⅱ): hypoxia/reoxygeuation, Group (ⅲ): hypoxia / reoxygenation + CT-1, Group (iv) : CT- 1 + hypoxia/ reoxygenation + LY294002 (PIK3/Akt inhibitor), Group (ⅴ): CT-1 + hypoxia / reoxygenation +DMSO. The concentration of CT-1 was 10 ng/mL. Myocytes survival rote was evaluated by MTS method, apopto-sis, mitochondrial permeability transition pore (△ψm) and reactive oxygen species(ROS) were detected by flow cy-tometer, phosphorylased GSK-3β and PI3K protein by western blotting. Analysis of variance and q test as statistical methods was used to analyze the data. Results Cardiomyocyte apoptosis and ROS increased markedly after hy-poxia/reoxygenation,but cardiomyocyte survival rate and the level of △ψm [(40.55±4.25) vs. (86.28±7.15), P < 0.01]decreased significantly. With CT-1 intervention, cardiomyocyte survival rate increased markedly (87%),apoptosis and ROS reduced significantly. The level of △ψm increased, the level of phosphorylased GSK-3β and phosphorylased PI3K protein obviously increased. The effect of CT-1 was inhibited by LY294002, but no significant effect was observed on ceils survival in DMSO group, which confirmed that LY294002 specifically in-volved blocking the protective effect of CT-1. Conclusions CT-1 can protect cardiac cells against hypoxia- reoxy-genation injury, these effects are dependent upon its ability to activate the PI3K/GSK-3β pathway.