Objective To detect the changes and the clinical significance in plasma protein-complement factor H-related protein 2 (CFHR2) in pulmonary arterial hypertension (PAH) associated with congenital heart disease (CHD) children.Methods Various types of 66 CHD patients with or without PAH and 16 healthy children(healthy control group) were studied,including 11 ventricular septal defects (VSD) with PAH (VSD-PAH),11 isolated VSD,11 atrial septal defects with PAH (ASD-PAH),11 isolated ASD,11 mixed type of heart defects [two or more defects of VSD,ASD and patent ductus arteriosus (PDA)] with PAH (Mix-PAH) and 11 cases without PAH (Mix).CFHR2 was validated by enzyme linked immunosorbent assay in the sample plasma.Results Compared with the healthy control group,the CFHR2 concentration in VSD-PAH patients [(189.10 ±24.01) μg/L vs.(42.99 ±4.53) μg/L,t =4.975,P ＜0.01] and VSD patients [(189.10 ±24.01) μg/L vs.(165.00 ±23.17) μg/L,t =2.661,P ＜ 0.05] were lower.The CFHR2 protein was also confirmed to be decreasing significantly in VSD-PAH patients compared with VSD patients (t =4.698,P ＜ 0.01).The plasma CFHR2 level in ASD-PAH patients [(189.10 ± 24.01)μg/L vs.(70.92 ± 8.27) μg/L,t =3.951,P ＜0.01] and ASD patients [(189.10 ±24.01) μg/L vs.(72.48 ± 8.99) μg/L,t =3.880,P ＜ 0.01] were significantly lower than those in the healthy control group,although there was no significant difference between ASD-PAH and ASD patients (t =0.128,P ＞ 0.05).The plasma CFHR2 level in Mix-PAH patients [(189.10 ± 24.01) μg/L vs.(83.23 ± 15.96) μL,t =3.314,P ＜ 0.05] was significantly lower than that in the healthy control group,while Mix patients [(189.10 ±24.01) μg/L vs.(170.40 ±33.15) μg/L,t =0.468,P ＞ 0.05] had no difference compared with the healthy control group,but had statistical significance with M ix PAH group (t =2.370,P ＜ 0.05).Conclusions The decrease of CFHR2 protein may demonstrate the deficiency of the immune system and coagulation mechanism in these patients and can be consi-dered as biomarker of CHD-PAH disease.

Since initiation of training on laboratory of molecular biology in my school, we accumulated experience in teaching process especially among international students and application of pedagogy as well as technology. Strengths, weakness and opportunities are analyzed to improve teaching quality.

BACKGROUND: Rational exercise training promotes bone and lipid metabolisms, while over-trained exercise makes negative effect. There are many biomarkers and pathways in the progress of bone and lipid metabolisms, which have been explored in different suitable studies. OBJECTIVE: To summarize the indexes and pathways of bone and lipid metabolisms, especially the common biomarkers and pathways, and to explore the effects of exercise on bone and lipid metabolisms. METHODS: PubMed and CNKI databases were retrieved with the keywords of "exercise training, bone metabolism, lipid metabolism, index, pathway" in English and Chinese, respectively. Finally, 52 eligible articles were enrolled for result analysis. RESULTS AND CONCLUSION: Bone metabolic indexes include 25-hydroxy-vitamin-D, calcitonin, parathyroid hormone, tumor necrosis factor, interleukin 1, 2, 4, 6 and 11, insulin-like growth factor 1, transforming growth factor-beta, bone morphogenetic protein, interferon, macrophage colony-stimulating factor, fibroblast growth factor, platelet fireworks growth factor, tartrate-resistant acid phosphatase, collagen Ⅰ telopeptide, pyridinoline, urinary deoxypyridinoline, alkaline phosphatase, osteocalcin, osteoporogeterin, carboxyterminal propeptide of type Ⅰ procollagen and so on. While lipid metabolic indexes contain triacylglycerol, serum total cholesterol, high density lipoprotein, low density lipoprotein, leptin, adiponectin, apolipoprotein, oxidized low density lipoprotein, and low density lipoprotein receptor related protein. Exercise intensity and duration are important factors for bone and lipid metabolisms. While a long-term high-intensity exercise will do harm to bone and lipid metabolisms, and a short-time high-intensity exercise is beneficial for bone metabolism. The interaction of exercise intensity and duration on bone and lipid metabolisms still needs to be further studied.

Objective To approach the evaluative effect of respiratory variation of superior vena cava peak flow velocity measured using transthoracic echocardiography (TTE) on fluid responsiveness in patients with mechanical ventilation.Methods A prospective cohort study was conducted.All mechanical ventilated critically ill patients whose fluid therapy was planned due to hypovolemia in Department of Critical Care Medicine of Beijing Tongren Hospital of Capital Medical University from April 2011 to April 2013 were enrolled.Volume expansion was performed with 500 mL Linger solution within 30 minutes.Patients were classified as responders if pulse pressure variation (PPV) increased ≥ 13％ before volume expansion.The respiratory variation in superior vena cava peak velocity was calculated as the difference between maximum and minimum values of velocity in peak A,peak S and peak D over a single respiratory circle,and their variations (ΔA,ΔS,ΔD) were also calculated.The receiver operating characteristic curve (ROC curve) was plotted to assess the evaluative effect of respiratory variation of superior vena cava peak velocity on fluid responsiveness.Results Twenty-seven patients were enrolled in this study.Volume expansion increased PPV ≥ 13％ happened in 14 patients (responders).The velocity of superior vena cava in peak A,peak S,peak D was significantly increased after volume expansion compared with that before volume expansion in responders [peak A (cm/s):34.6 ± 2.2 vs.31.3 ±2.1,t=-2.493,P=0.027; peak S (cm/s):39.1 ± 1.3 vs.35.3 ±2.1,t=-2.564,P=0.024; peak D (cm/s):28.1 ± 1.2 vs.23.3 ± 1.4,t=-4.995,P=0.000],but there was no significant difference in ΔA,ΔS and ΔD between before and after volume expansion.The ΔA,ΔS and ΔD were positively correlated with PPV (r＝0.040,P=0.854; r=0.350,P=0.074; r=0.749,P=0.000).The area under ROC curve (AUC) of peak S was 0.36 [95％ confidence interval (95％CI):0.11-0.52],but the AUC of ΔS was 0.68 (95％CI 0.47-0.89),the AUC of peak D was 0.41 (95％CI 0.19-0.63),but the AUC of ΔD was 0.95 (95％CI 0.86-1.00),so the aberration rate of superior vena cava in respiration was better than the flow rate in superior vena cava.When the cut-off value of ΔS was 20.7％ for predicting fluid responsiveness,the sensitivity was 78.6％ and the specificity was 61.5％.When the cut-off value of ΔD was 12.7％ for predicting fluid responsiveness,the sensitivity was 92.0％ and the specificity was 92.3％.Conclusion Respiratory variations in superior vena cava peak velocity measured by TTE could assess fluid responsiveness in patients with mechanical ventilation.

BACKGROUNDTo date murine models of permanent focal cerebral ischemia have not been well characterized. The purposes of this paper were to compare three different permanent middle cerebral artery occlusion (MCAo) models with or without craniectomy, and to identify an ideal mouse model of permanent focal cerebral ischemia.

METHODSExperiments were performed on 45 healthy adult male Kunming mice, weighing 28 to 42 g. The animals were randomly assigned to three groups (n = 15 in every group) based on surgical procedure: MCAo via the external carotid artery (ECA), MCAo via the common carotid artery (CCA), and direct ligation of the middle cerebral artery (MCA). Each day post-ischemia, the animals were scored using an eight-grade neurological function scale, and mortality was also recorded. Seven days post-ischemia, the brains were removed for lesion size determination using triphenyltetrazolium chloride staining. Correlation analysis of lesion volume and neurological score was carried out.

RESULTSMortality in the group receiving direct MCA ligation was lowest among the three groups, and there was a significant difference between the direct MCA ligation group and the two intraluminal occlusion groups (P < 0.05). In all groups, neurological scores gradually increased with prolongation of ischemic duration, peaking after two days, then gradually decreasing. In the direct MCA ligation group, however, neurological scores were relatively stable. There was a significant correlation between infarct volume and neurological score 7 days after MCAo in every group (all r > 0.7, P < 0.05), suggesting good reproducibility of lesion volume in the three groups, but the infarct volume was more constant in the direct MCA ligation group.

CONCLUSIONThe direct ligation model of MCAo provides an optimal means of studying permanent focal cerebral ischemia, and is preferable to the models using intraluminal sutures.

Animals ; Brain Ischemia ; Disease Models, Animal ; Ligation ; Male ; Mice ; Middle Cerebral Artery ; surgery ; Random Allocation ; Reproducibility of Results

Objective To investigate the effect of platelet-rich plasma(PRP) combined with bone graft in the treatment of humeral condylar bone defect.Methods A total of 135 patients with humeral condylar bone defect in Ankang central hospital from January 2012 to December 2015 were divided into the PRP combined group(n =69) and the conventional group(n =66) according to the order of admission time.The patients of PRP combined group were treated with platelet-rich plasma combined with autologous bone graft,and patients of conventional group received autologous bone graft,respectively.The surgery time,hospitalization time,wound healing,fracture union and the motion of elbow joint at postoperative 1 year between two groups were compared.The Kaplan-Meier survival curve was used to reflect the bone healing in both groups,and the log-rank test was used to compare the result.Results There was no statistically significant difference in the surgery time,hospitalization time,wound healing and motion of elbow joint at postoperative 1 year between the two groups(P ＞ 0.05).But the average time of wound healing (3.8 ± 0.72) weeks and the time of bone union (18.8 ± 3.50) weeks in PRP combined group were significantly shorter than (6.4 ±0.58) weeks and (22.7 ± 1.55) weeks in the conventional group(P =0.000),the differences were significant.The KaplanMeier survival curve of the bone union in the PRP combined group was also significantly better than that in the conventional group.Conclusion PRP can promote the healing of fracture in patients with humeral condylar bone defect after autologous bone graft,which contributes to the recovery of elbow function.

Objective This study aims to elucidate the protective role of Acacetin against apoptosis in HP-infected GES-1 cells and to delineate its potential underlying mechanisms.Materials and Methods GES-1 cells were subjected to in vitro treatment with HP and Acacetin.Cell viability was assessed utilizing the CCK-8 assay,alterations in cell migration and healing capacities through the wound healing assay,rates of apoptosis via flow cytometry,and expression levels of apoptosis-associated proteins through western blot analysis.Results HP infection led to a diminution in GES-1 cell viability,a suppression of cell migration,an augmentation in the rate of apoptosis,and an increase in the expression levels of Bax and cle-caspase3.Conversely,treatment with Acacetin was found to enhance cell viability,mitigate apoptosis induced by HP infection,and modulate the expression of apoptosis proteins by downregulating Bax and cle-caspase3.Discussion and Conclusion Acacetin significantly improves GES-1 cell vitality and inhibits apoptosis in HP-infected GES-1 cells,thereby offering a protective effect on gastric mucosal epithelial cells.

After treating with chemotherapy or immunosuppressant, malignant diseases of hematopoietic system such as leukemia, malignant lymphoma and aplastic anemia usually induced severe infection such as sepsis. Sepsis which is hard to be diagnosed causes high death rate. This study was purposed to establish an experimental sepsis mouse model so as to provide a basis for pathogenesis and intervention study. A classic caecal ligation and puncture (CLP) was used to establish experimental sepsis model. ELISA was used to detect levels of C5a, IL-6, TNFalpha, and IFN-gamma. Flow Cytometry was applied to measure apoptosis of lymphocytes in thymus and mesentery. The pathologic changes of thymus and spleen were confirmed by HE staining. The results showed that almost 70%-80% mice died at 72 hours after CLP. Only approximate 20% animal survived during finite time, mice in CLP group had significant weight lose. Meanwhile large release of different inflammatory mediators which are related with sepsis (C5a, IL-6, TNF-alpha, and IFN-gamma) was observed after CLP. Apoptosis of lymphocytes in thymus and mesentery lymphonodus was enhanced markedly after CLP. Significantly pathologic injury was also observed in thymus and spleen. It is concluded that a mouse model of experimental sepsis was successfully established by caecal ligation and puncture which can well mimic the clinical symptom of sepsis. The experimental sepsis mouse model provides an excellent tool for exploring the pathogenesis and intervention ways for sepsis accompanied with complicated malignant hematological diseases in vivo.
Animals ; Apoptosis ; Cecum ; injuries ; Complement C5a ; metabolism ; Disease Models, Animal ; Interferon-gamma ; metabolism ; Interleukin-6 ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Sepsis ; metabolism ; pathology ; Spleen ; pathology ; Thymus Gland ; pathology ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo establish a bone metastasis model of prostate cancer by intratibia injection of Du145 in nude mice, observe the local growth of tumor in tibia and then assess application value of this model.

METHODSFor 9 male nude mice, Du145 (5 x 10(6)) was injected in tibia by a TB syringe with a 29-gauge needle at a dose of 30 microl per mouse. Then the vital signs of the nude mice were observed. When the mice were dying, they were sacrificed, and the tissues of right hindlimbs, lymphatic nodes, lungs and livers were taken out, fixed in 10% formalin, embedded in paraffin, stained by HE and then observed microscopically.

RESULTSIncidence of bone tumor after intratibia injection was 67% (6 out of 9). About 48 days later, there were some small palpable nodes in right hind-limbs of the 6 mice and they couldn't walk normally. About 55 days later, cachexia occurred in them. After dissection, some carrion-like tissue grew from marrow cavity to muscular spatium, which was identified as tumor tissue by HE. The envelop of livers became crampy, and acute hepatitis could be diagnosed through microscopy, which represented a large scale of hepatocytic death, liver sinus dilatation and hyperemia, hepatic lobule infiltrated by lymphocyte, macrophage and inconspicuous hyperplasia. Since hypohepatia occurred too early, we couldn't detected distant metastases.

CONCLUSIONThe intratibia injection model is an optimal animal model to study metastasis of prostate cancer. It mimics the natural situation of human prostate cancer and will help to understand the mechanisms of androgen-independence and osseous metastasis, and tumor-host determinants of PSA expression.

Animals ; Bone Neoplasms ; secondary ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Prostatic Neoplasms ; pathology ; Tumor Cells, Cultured ; Xenograft Model Antitumor Assays ; methods

OBJECTIVETo construct a lamivudine-resistant plasmid containing 1.2 unit genome of duck hepatitis B virus and identify its replication and drug-resistance in avian LMH hepatica cells.

METHODSThe recombinant plasmid PBS-DHBV1.2 was constructed using the 1.2-genome length DHBV DNA sequence from a dimer DHBV genome with pcDNA3.1 as the template. With site-directed mutagenesis, we obtained PBS-DHBV1.2-M512V plasmids with polymerase gene mutation from PBS-DHBV1.2. Two constructed plasmids were transiently transfected into LMH cells using FuGENETM6 transfection reagent and cultured in the medium containing different concentrations of lamivudine. Southern blot hybridization was performed to detect DHBV replication intermediates.

RESULTSPCR amplification, restriction digestion and plasmid sequencing all confirmed successful construction of PBS-DHBV1.2-M512V recombinant plasmid. Southern blot analysis identified the presence of all the expected DHBV replication intermediates in LMH cells. The replication capacity of the mutant plasmid was decreased by 2.7 times compared with that of the wild plasmid. The IC(50) of lamivudine was 37.12∓8.81 ng/ml for the mutant, greater than that of the wild plasmid (10.90∓4.80 ng/ml).

CONCLUSIONCompared with the wild plasmid, the mutant plasmid has a lower replication capacity and sensitivity to lamivudine in vitro.

Antiviral Agents ; pharmacology ; Drug Resistance, Viral ; drug effects ; genetics ; Hepatitis B Virus, Duck ; drug effects ; genetics ; Lamivudine ; pharmacology ; Mutagenesis, Site-Directed ; Plasmids