Abstract:Objective To establish a method for isolating and purifying tumor-associated hepatic stellate cells (tHSCs) from human hepatocellular carcinoma (HCC) tissues and to study their impact on the biological behaviors of HCC cells in vitro. Methods The tHSCs were isolated from the fresh human HCC tissue by collagenase/pronase digestion, followed by density gradient centrifugation using Nycodenze. The viability of the isolated cells was determined by trypan blue staining assay. The tHSCs were identified by immunocytochemical staining of α-SMA, a specific marker for activated hepatic stellate cells. The morphologic changes of the tHSCs were observed under microscope. The conditioned medium (CM) of tHSCs cultured in serum-free DMEM was collected, and then used for co-culture with HCC cell lines QGY-7701, BEL-7402, and SMMC-7721. CM of human hepatic stellate cell line LX-2 was used as control. Cell viability, proliferation, migration potential and EMT-related protein expression changes were assessed by using the MTT assay, wound migration assay and Western blotting analysis, respectively. Results The viability of isolated tHSCs was over 95%, with a purity about 100%, and the isolated tHSCs could be passaged and cryopreserved. tHSCs significantly induced typical EMT-like morphological changes and altered expression of molecular markers in the three HCC cell lines, with the epithelial marker E-cadherin significantly decreased (P<0.05) and the stromal marker Vimentin significantly increased (P<0.05). Meanwhile, tHSCs efficiently enhanced the proliferation and migration of HCC cells in vitro. Conclusion We have established a method for isolating and purifying tHSCs from human HCC tissues, which can help future study on tHSCs. tHSCs can enhance the proliferation and migration of HCC cells through inducing EMT-like phenotype.

Abstract:Objective To investigate the effects of type-3-polysaccharide (T3P) on airway inflammation and Treg/Th17 cells in bronchial asthmatic mice, so as to explore the immunoregulation mechanism of T3P in asthmatic mice. Methods BALB/c mice were randomly divided into 3 groups, with 8 mice in each group. The control group received PBS treatment, the asthma group were sensitized and challenged with ovalbumin(OVA),and the T3P group received a pretreatment with T3P by subcutaneous injection before sensitization with OVA. The pulmonary histological changes were observed and differential cell counts in bronchoalveolar lavage fluid(BALF) were performed. Serum OVA-IgE and IFN-γ, IL-4, IL-17 and IL-10 levels in the BALF were detected by enzyme-linked immunosorbent assay (ELISA). The levels of Foxp3 and RORγt mRNA expression were measured by real-time fluorescence-based quantitative PCR. The proportions of Treg and Th17 cells in CD4⁺ cells were assessed by flow cytometric analysis. Results The inflammatory degree of pulmonary tissue, serum OVA-IgE, and total cell number, proportion of eosinophils, and IL-4, IL-17 levels in BALF were significantly lower in T3P group than in the asthma group (P<0.05). The BALF levels of IFN-γ and IL-10, Foxp3 mRNA expression and proportion of Treg cells among CD4⁺ cells were significantly higher in the T3P group than in the asthma group (P<0.05), while RORγt mRNA expression and the proportion of Th17 cells among CD4⁺ cells were significantly lower than in the asthma group (P<0.05). The count of BALF eosinophils was negatively correlated with the ratios of Foxp3-mRNA/RORγt-mRNA and Treg/Th17 in asthma and T3P groups (P<0.05). Conclusion T3P may inhibit airway inflammation by inhibiting OVA-IgE and regulating Th1/Th2 and Treg/Th17 cell balance in asthmatic mouse model．

Abstract:Objective To observe C4d deposition in renal allografts of rats undergoing chronic allograft nephropathy (CAN), and to analyze the effects of immunosuppressants on deposition of C4d in peritubular capillaries. Methods The renal grafts of Fisher 344 rats were orthotopically transplanted into Lewis rats to create CAN models, and all the recipients were given cyclosporine A (CsA) 10 mg/(kg· d)×10 d after operation. The models were then divided into 5 groups (each n=9): Group A was normal saline control group, only receiving vehicle orally; Group B, C, D, and E received CsA 6 mg/(kg·d), RAPA 0.8 mg/(kg·d),FK506 0.15 mg/(kg·d),and MMF 20 mg/(kg· d), respectively. The renal allografts were harvested after three rats were sacrificed at the 4^th, 8^th and 12^th weeks post-transplantation. The histological changes were assessed according to Banff 97 standard. The deposition of C4d was detected by immunofluorescence method. Results C4d deposition in peritubular capillary(PTC) was found in all the allografts at the 4^th week after transplantation, while there were no obvious clinical pathological changes of CAN in all groups, and the Banff scores were not significantly different among different groups (P>0.05). CAN manifestations of different degrees were observed 8 weeks after operation, with increased C4d deposition in the PTC. Severest CAN was observed at the 12^th week after operation, accompanied by the most C4d deposition in the PTC. C4d deposition was positively correlated with the severity of CAN (r=0.894, P=0.000). Compared with the control group, CsA and FK506 showed no significant effect on C4d deposition (P>0.05); however, MMF and RAPA significantly decreased C4d deposition (P<0.05). Conclusion Deposition of C4d in PTC may appear in allografts earlier than the pathological changes of CAN, and the deposition is associated with the progression of CAN. MMF and RAPA can inhibit the progression of CAN, while CsA and FK506 can not.

Abstract:Objective To establish a New Zealand rabbit bone marrow mesenchymal stem cells (BMSCs)-gentamicin-calcium alginate 3D sustained-release culture system and to study the growth and differentiation of BMSCs. Methods BMSCs-calcium alginate 3D culture system (W group) and BMSCs-gentamicin-calcium alginate 3D sustained-release culture system (U group) were constructed and were cultured with HG-DMEM (15% FBS, 10 ng/mL TGF-β₁) under saturated humidity, 37℃ and at 5% CO₂ , with the culture medium changed on a daily basis, and the cell morphology and microsphere morphology changes were observed. H-E staining, toluidine blue staining and type Ⅱ collagen staining were performed for the microspheres on week 2, 4, and 6. Results Cell clusters were formed locally in the two groups after the 3D microspheres were cultured for 10 days. A large number of cell clusters were formed after 21 days, and BMSCs maintained a spherical or approximate spherical shape. There was no significant difference in cell proliferation or growth between the two groups (P>0.05). After a 2-week culture, toluidine blue staining of microspheres showed positive staining in both groups, but with no obvious extracellular matrix formation, and staining for collagen type Ⅱ antibody was weakly positive. After a 4-week culture, toluidine blue staining was obvious in the periphery of the cell microspheres in both groups, but the staining was unapparent in the center; extracellular matrix around the cell clusters had less blue colored substance, and the central cell clusters had more mauve substance; collagen type Ⅱ staining was strongly positive in both groups. Conclusion Local sustained-release of appropriate amount of gentamicin has no noticeable effect on the growth and transformation of BMSCs while reaching the minimum inhibitory concentration. The influence of Gentamicin on ultrastructure of BMSCs and chondrocytes remains to be further investigated.

Abstract:Objective To primary culture human aortic valvular interstitial cells (hVICs), establish their in vitro calcification model, and to induce hVICs differentiation to osteogenesis and to observe the phenotype changes. Methods hVICs were digested from native valves and used for experiments after 3-7 passages. The cells were cultured in osteogenic media or in normal media. One week later the calcified nodules were stained and measured by von Kossa. The activity of alkaline phosphatase (ALP) was examined by spectrophotometer, immunofluorescence staining was used to detect the phenotype protein of hVICs, and real-time PCR and Western blotting analysis were used to examine the osteogenesis associated factors to assess the calcification model of hVICs. Results The calcified nodules were found 7 days after osteogenic induction. The calcified nodules in the experimental group were significantly more than that in the control group (51.20±14.31/well vs 3.60±1.82/well,P<0.05). The activity of ALP was significantly increased after osteogenic induction compared with the control group (increased by about 4 folds, P<0.05), with increased contractile phenotype α-smooth muscle actin (α-SMA). Real-time PCR and Western blotting results indicated that the expressions of Runx2, osteocalcin, and osteopontin in the experiment group were significantly higher than those in the control group (P<0.05), so was the expression of phosphorylated Smad1/5/8(P<0.05). Conclusion We have successfully established the in vitro calcification model of hVICs, with hVICs in an activated state; and the phenotype shifts to contraction and ossification, which provide a reliable cell model for the future study.

Abstract:Objective To detect subtle mutations in the dystrophin gene by exome capturing using second-generation sequencing technique. Methods Exome capturing using second-generation sequencing technique was used to detect mutations of dystrophin gene in a patient with typical clinical manifestations of Duchenne muscular dystrophy （DMD）, but without deletions or duplications in the dystrophin gene. The mutations were verified by Sanger sequencing, and bioinformatics was employed to predict its influence on the coding. The patient’s mother and 100 healthy volunteers were taken as controls.Results A base change in the first base of intron 50 (G>C) was found in dystrophin gene of the patient, and his mother was heterozygosis at the same site. Bioinformatics predicted that the 5′ donor splicing site of intron 50 would disappear due to this base change, which would alter the amino acid sequence at the C terminal of corresponding peptide and result in the appearance of premature termination codon. Sanger sequencing confirmed that the base change was a novel pathogenic mutation in the dystrophin gene, and it was absent in normal controls. Conclusion It is demonstrated that exome sequencing technique can effectively detect the subtle mutations in the dystrophin gene, which may contribute to better molecular diagnosis of DMD.

Abstract:Objective To observe the therapeutic effect of Bushen huoxue huatan recipe (BHHR) on androgen-induced sterilized rats (ASR) and the expression of androgen synthase and metabolic enzymes in the ovary of ASR before and after treatment with BHHR, and to discuss the possible mechanism. Methods Female SD rats of 9-day old were injected subcutaneously with testosterone propionate (1.25 mg) to create model. The model rats were randomly divided into 3 groups: model group (treated with distilled water by gastrogavage, 10 mL/kg), metformin therapy group (gastrogavage, 0.1 g/kg) and BHHR therapy group (gastrogavage, 10 mL/kg), with 13 animals in each group. Ten rats with normal estrous cycle served as normal controls. The body mass, sexual cycle recovery and ovary mass/body mass ratio were observed after 28-day treatment. Serum testosterone level was measured by radioimmunoassay; the expressions of 3β-hydroxylsteroid dehyrogenase (3β-HSD), cytochrome P450 17α-hydroxylase/17, 20-lyase (CYP17) and P450 aromatase (P450arom) in ovary were detected by quantitative immunohistochemistry method. Results (1) Significantly more rats in the metformin and BHHR groups had sexual cycle recovery compared with that in the model group (P<0.01). (2) The body mass of rats in the model group was significantly heavier than that in the normal control group (P<0.01), and those of the BHHR and metformin groups were significantly lighter than that of ASR model group (P<0.01). (3) Ovary mass/body mass ratio of the model rats was significantly higher than those of the other three groups (P<0.01). (4) Serum testosterone level in the model group was significantly higher than those in the other three groups (P<0.01). (5) Compared with the model group and the normal group, the P450arom expression in the BHHR group was significantly increased (P<0.01), while no significant difference was found in 3β-HSD and CYP17 expression between the BHHR group and the model group (P>0.05). Conclusion BHHR can reduce serum testosterone levels in ASR rat, which might be through up-regulating the expression of the androgen metabolism enzyme P450arom.

Abstract:Objective To explore the effect of MPT64 antigen from Mycobacterium tuberculosis on RAW264.7 macrophages and the related mechanism. Methods MPT64 was purified after expression in E.coli and verified by Western blotting analysis. The RAW264.7 differentiation was induced by phorbol myristate acetate (PMA) and the resultant cells were divided into three groups according to different treatments: negative control, purified protein derivative of BCG(BCG-PPD) and BCG-PPD+MPT64 treatment groups. After 16 h incubation, flow cytometry was used to examine apoptosis of macrophages, and the levels of TNF-α and IL-10 in the supernatants were determined by ELISA. Results BCG-PPD treatment induced apoptosis of RAW264.7 macrophages, and compared with BCG-PPD group, the apoptotic level of macrophages was significantly lower in BCG-PPD+MPT64 group (P<0.05). We also found that the supernatant TNF-α level in the BCG-PPD group was significantly higher than that in negative group (P<0.01) and the IL-10 levels were not significantly defferent between the two groups. Compared with BCG-PPD group, the IL-10 level was significantly increased in BCG-PPD+MPT64 group (P<0.01) and the TNF-α levels were not significantly different between the two groups. Conclusion MPT64 may act as a virulence factor and can inhibit the apoptosis of macrophages induced by BCG-PPD, which is probably through increasing IL-10 level.

Abstract:Objective To explore the clinical characteristics and risk factors of newly diagnosed type 2 diabetic mellitus (T2DM) patients and to investigate the correlations among nonalcoholic fatty liver disease (NAFLD), hyperuricemia, neck circumference (NC) and carotid intima-media thickness (CIMT) in these patients. Methods The 85 patients were all within one year after diagnosis and they were hospitalized immediately after diagnosis.The clinical data including NC, waist circumference (WC), waist-hip ratio (WHR), and body mass index (BMI) were collected.The fasting insulin, fasting C-peptide, hemoglobin A_1c, liver and renal function, phenotype of lipid and uric microalbumin were determined, and ultrasound examination of the liver and carotid artery was performed. Results The prevalence rates of NAFLD, hyperuricemia and increased CIMT were 57.6% (49/85), 16.5% (14/85) and 29.4% (25/85), respectively.Up to 88.2% (75/85) of the patients had two or more metabolic abnormalities. Patients with NAFLD, hyperuricemia and increased NC had significantly more metabolic abnormalities compared with those without the corresponding conditions (P<0.05). The BMI, NC, WC, alanine aminotransferase (ALT), and triglyceride (TG) were significantly different among different groups (all P < 0.05).Multivariate unconditional logistic regression analysis showed that NC, WC, BMI, ALT, fasting C-peptide, and serum uric acid (SUA) were common risk factors of NAFLD and increased CIMT. Conclusion Most newly diagnosed T2DM are concomitant with a variety of metabolic abnormalities.NAFLD, hyperuricemia, increased NC and CIMT are risk factors to each other.NC and SUA are also the common risk factors of NAFLD and increased CIMT.

Abstract:Objective To investigate the relationship between the clinical factors of renal transplant recipients medicated with cyclosporine A (CsA) and gingival overgrowth (GO), so as to evaluate the association of serum cyclophilin A (CyPA) with CsA-induced GO. Methods Totally 65 renal transplant recipients were enrolled in this study. The degrees of GO were evaluated using gingival overgrowth index (GOI) and the patients were subsequently grouped into overgrowth (GO+) and non-overgrowth (GO-). The age, gender, medication duration, dosage of immunosuppressive agents, serum concentrations of CsA and CyPA, creatinine, ureal and gingival assessments were compared between the two groups. The possible correlation between CyPA level and the development and severity of CsA-induced GO was also identified. Results The number of GO+ subjects (GOI≥30) was 14 (21.54%) in the present patients. There were no significant differences in age, gender and medication duration, dosage of immunosuppressive agents, serum concentration of CsA, serum creatinine or ureal level between the two groups. Serum CyPA level in GO+ group was significantly lower than that in GO- group (0.23 \[0.16-0.30\] ng/mL vs 0.34\[0.22-0.54\] ng/mL, P=0.04). Serum CyPA concentration was negatively correlated with GO degree (r=-0.264, P=0.03), and was not correlated with dosage or concentration of CsA (r=-0.014, P=0.91; r=0.012, P=0.93). In addition, the patients with GO presented a significantly high plaque index and papilla bleeding index than those without GO (1.41±0.27 vs 1.15±0.34, P=0.01; 0.49±0.30 vs 0.25±0.11, P=0.01). Conclusion Development of CsA-induced GO may be due to a co-work of both local and system factors; serum CyPA level might be an risk factor for CsA-induced GO independent to dosage and serum level of CsA, which may help to detect GO in clinic.

Abstract:Objective To evaluate the feasibility, safety and efficacy of Solitaire stent placement for treatment of complex intracranial atherosclerotic stenosis. Methods We retrospectively reviewed the clinical data of 18 patients who underwent Solitaire stent placement for symptomatic intracranial arterial stenoses in our department between November 2011 and June 2012, with focus on the stenosis morphology, condition of the diseased blood vessels, surgical success rate, and perioperative complications. We also summarized the early outcomes and imaging findings during follow-up. Results The patients included 8 males and 10 females, with a mean age of 66.55±9.84 years old. Mori typing results were 5 A, 8 B, and 5 C types; LMA typing results were good in 2, moderate tortuosity in 12, severe tortuosity in 4 cases. All the patients received successful Solitaire stent implantation. The average stenosis rate was reduced from (74±12.5)％ to (10±5.5)% after the procedure. There was only one patient (5.6%) had post-operative complication (cerebral infarction of the target vessel territory)，who had temporal symptom and recovered well at follow-up.Clinical follow-up was available for 18 patients, with a mean of 6.2± 1.4 months; and no patients experienced any ischemic episodes. Fifteen patients had DSA or CTA follow-up for a mean of 7.8±2.6 months. There were 3 in-stent restenosis (20%) cases and one of them was symptomatic with transient ischemic attack. Conclusion Solitaire stent placement is feasible, safe and effective for complex symptomatic intracranial artery stenosis, with definite short-term effectiveness, and the long-term effect warrants further investigation.

Abstract:Objective To assess the surgical outcomes of double-level isthmic spondylolisthesis in adults. Methods From Mar.2006 to Nov.2010, 30 patients with double-level lumbar isthmic spondylolisthesis were treated with modified Jaslow technique combined with posterior lumbar interbody fusion. Clinical outcomes were assessed by Japanese Orthopedic Association (JOA) score; the functional disability was quantified by Oswestry Disability Index (ODI); the Lenke grading system was used to assess the spinal fusion; and the Henderson grading system, the intervertebral height index, and the Taillard index were used to assess the clinical outcome. Results All the 30 patients achieved reduction of different degrees, with the mean reduction rate being (43.60±25.29)%. The intervertebral height index was (17.68±6.67)% before operation, increased to (50.99±4.94)% at 1 week after operation and (44.37±5.84)% at the last follow-up. The Taillard index was (20.91±7.11)% before operation, (10.23±2.16)% at 1 week after operation, and (11.87±2.73)% at the last follow-up. The height index and the Taillard index before operation were significantly different from those of the other two time points (1 week after operation and final follow-up, P<0.01). The bone fusion was grade A in 49 levels and grade B in 11 levels. The JOA score increased from 9.60±1.89 before operation to 22.40±2.06 at 6 months after operation and 22.70±1.63 at the final follow-up. ODI was 34.90±5.56 before operation, which was decreased to 11.80±1.81 at 6 months after operation and 10.70±1.33 at the last follow-up. The JOA score and the ODI index before operation were significantly different from those of 6 months after operation and those at the final follow-up (P<0.01). The Henderson clinical outcome was excellent in 19 cases, good in 8 cases, and poor in 3 cases. Conclusion Complete decompression, reduction of isthmic spondylolisthesis by modified Jaslow technique, pedicle screw fixation, and posterior lumbar interbody fusion can achieve satisfactory clinical results for adults with double-level isthmic spondylolisthesis.

Abstract:Objective To investigate the expression of interleukin (IL)-12 and IL-17 in the serum and liver tissues of patients with autoimmune liver disease (AILD) and to discuss the relevant significance. Methods The peripheral blood and liver tissues were collected from 21 patients with autoimmune hepatitis (AIH), 21 with primary biliary cirrhosis (PBC), and 9 with AIH-PBC overlap syndrome (AIH-PBC OS). The serum IL-12 and IL-17 levels were assayed by ELISA, and the expression and location of IL-12 and IL-17 in the liver tissues were detected by immunohistochemistry method. Alamine transaminase(ALT) and γ-glutamyltransferase(GGT) were measured by automatic biochemical analyzer. The correlation of serum IL-12 and IL-17 expressions with ALT and GGT levels was analyzed. Ten healthy participants taking a physical examination and 10 normal liver tissues were used as controls. Results The serum levels of IL-12 in AIH, PBC, and AIH-PBC OS groups were significantly lower than that in the control group(P＜0.01), and the serum levels of IL-17 in the three groups were significantly higher than that in the control group(P＜0.01). A negative correlation was found between serum IL-12 and IL-17 in AIH and PBC groups (AIH: r=-0.752,P＜0.05；PBC:r=-0.436, P＜0.05), and serum IL-17 was positively correlated with ALT and GGT (AIH: r=0.825, P＜0.05；PBC:r=0.571, P＜0.05). IL-12 was mainly expressed in bile duct epithelium and Kupffer cells, and the expression rates in the 3 experimental groups were significantly lower than that in the control group (AIH: 19.05% \[4/21\], PBC: 9.52% \[2/21\], AIH-PBC OS: 11.11% \[1/9\], control: 90.00% \[9/10\]; P＜0.01). IL-17 was mainly expressed in lymphocytes and monocytes, and the expression rates in the 3 experimental groups were significantly higher than that in control group (AIH: 71.43% \[15/21\], PBC: 76.19% \[16/21\], AIH-PBC OS: 77.78% \[7/9\], control: 10.00% \[1/10\]; P＜0.01). IL-12 expression in the liver tissues was negatively correlated with IL-17 expression in AIH and PBC groups(AIH:r=-0.499, P=0.021; PBC: r=-0.580,P=0.006). Conclusion IL-12 expression is reduced in AILD patients, decreasing the inhibition against Th17 cell differentiation, leading to IL-17 increase and mediating inflammatory injury, which indicates that IL-12/IL-17 inflammatory pathway may participate in the development and progression of AILD.

Abstract:Objective To assess the performance of the European System for Cardiac Operative Risk Evaluation Ⅱ(EuroSCORE Ⅱ) in predicating in-hospital mortality among Chinese patients undergoing heart valve surgery at our center. Methods From January 2006 to December 2011, 3 479 consecutive patients who underwent heart valve surgery at our center were enrolled in this study and they were scored by the original EuroSCORE(addtive EuroSCORE and logistic EuroSCORE) and EuroSCORE Ⅱ model. The actual mortality rate of patients was compared with those of the predicted ones. The performances of the original EuroSCORE and EuroSCORE Ⅱ model were assessed by the Hosmer-Lemeshow (H-L) test. The discrimination validity of prediction was tested by calculating the area under the receiver operating characteristic (ROC) curve. Results There were 112 in-hospital deaths among the 3 479 patients, with an in-hospital mortality rate of 3.2%, compared to the predicted mortality rates of 3.84% by the additive EuroSCORE (H-L: P=0.013, suggesting a higher prediction) ,3.33% by the logistic EuroSCORE (H-L: P=0.08, suggesting good consistency), and 2.52% by the EuroSCORE Ⅱ (H-L: P<0.0001, suggesting a lower prediction).EuroSCORE Ⅱ showed a good calibration in predicting in-hospital mortality for patients undergoing single valve surgery (H-L: P=0.103, area under the ROC curve of 0.792) and a poor calibration for patients undergoing multiple valve surgery (H-L: P<0.0001, area under the ROC curve of 0.605). The discriminative powers of the predictions by additive EuroSCORE, logistic EuroSCORE, and EuroSCORE Ⅱ were poor for the entire cohort, with the areas under the ROC curve being 0.684, 0.673, and 0.685, respectively.Conclusion EuroSCORE Ⅱ has a better accuracy for predicting mortality of patients undergoing single valve surgery, but not for those undergoing multiple valve surgery, which should be considered in clinical practice.

Abstract:Objective To investigate the appearance of 3.0 T magnetic resonance imaging (MRI) of acute myocarditis (AM) and the related clinical significance. Methods A total of 25 patients with AM were assigned to myocarditis group and 15 healthy volunteers were assigned to control group, all cases underwent cardiac MRI enhancement. The protocol included dual inversion recovery T2-weight null fat, diffusion weight imaging (DWI), cine, first-pass perfusion and delayed enhancement MRI. Results No positive abnormality was found in the 15 healthy volunteers. In the myocarditis group, 4 (16.0%) cases showed focal spot or patchy high signal in the left ventricular (LV) myocardium in dual inversion recovery T2-weight null fat. DWI in LV myocardium showed hyperintensity in 6 (24.0%) cases. Cine revealed reduced local ventricular wall motion in 4 (16.0%) patients. The ejection fraction (EF), LV end-diastolic volume and LV end-diastolic diameter in the 25 AM patients were(58.9±4.7)%, (147.0±28.9) mL, and (51.8±4.7) mm, respectively, which were not significantly different from those of the control group (P>0.05). One (4.0%) case had reduced local myocardial perfusion during first-pass perfusion. A total of 53 myocardial segments were involved in the 23 (92.0%) cases with subepicardial or/and mid-myocardial delayed enhancement, which mainly located in the septal zone and lateral zone. Ventricular wall motion abnormalities were consistent with delayed enhanced segments. Conclusion 3.0 T MRI can display the position, shape, range, degree and function of AM. Comprehensive analysis of cardiac MR sequences can provide imaging evidence for diagnosis and differential diagnosis of AM.

Abstract:Objective To explore the association of body mass index (BMI) with high-resolution CT (HRCT) volumetric data and HRCT phenotypes in patients with chronic obstructive pulmonary disease (COPD). Methods Sixty-two consecutive COPD patients underwent HRCT and their weight and height were recorded. COPD subjects were classified into three phenotypes based on the visual HRCT findings (with or without emphysema and/or bronchial wall thickening). With the volumetric HRCT data, the total lung volume (TLV) was calculated automatically by Extended Brilliance Workspace^TM , the total emphysema volume (TEV) was obtained by applying density thresholds of -950 HU, and the TEV/TLV was calculated as an emphysema index (EI). The correlation between the volumetric HRCT data and BMI was assessed using Spearman correlation analysis. The three phenotypes of COPD subjects were evaluated using one-way ANOVA. Results Visual HRCT findings showed that the COPD patients were classified into 3 phenotypes: phenotype A (n=42), phenotype E (n=9) and phenotype M (n=11). BMI of COPD patients was correlated with both TEV (r=-0.389, P=0.002) and EI (r=-0.424, P=0.001). COPD patients with phenotype A had a significantly higher BMI (\[23.4±3.4\] kg/m²) compared with those with phenotype E (\[20.6±3.1\] kg/m², P<0.05). The BMI of patients with phenotype M(\[21.3±2.7\] kg/m² ) was not significantly different from those of phenotypes A or E. Conclusion The TEV and EI are negatively correlated with BMI in COPD patients. HRCT phenotype A compared with phenotype B and C is associated with a higher BMI in COPD patients.

Abstract:Objective To investigate the effect of recombinant human endostatin on the expression of vascular endothelial growth factor (VEGF) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in hypertrophic scar (HS) of rabbit ears. Methods A total of 16 New Zealand rabbits were used to establish ear HS models. The model rabbits were evenly randomized into two groups: experimental group and control group. Rabbits in the experimental group were locally injected with recombinant human endostatin (0.1 mL, 5 mg/mL, every other day for five times) and those in the control group were injected with normal saline (0.1 mL, every other day for five times). The HS specimens were harvested 30 days after intervention and were grossly observed; RT-PCR was used to test the expression of VEGF and TIMP-1 mRNA, and TIMP-1 protein was examined by immunohistochemistry method. Results The mRNA expression levels of VEGF and TIMP-1 in experimental group were significantly lower than that in control group (VEGF: 0.279 0±0.030 7 vs 0.657 0±0.161 1, P<0.05; TIMP-1: 0.244 4±0.057 4 vs 0.730 2±0.103 8, P<0.05). The protein expression level of TIMP-1 in experimental group was significantly lower than those in control group (P<0.05). Conclusion Recombinant human endostatin can inhibit HS of rabbit ear, which might be through suppressing the expression of VEGF and TIMP-1.

Abstract:The mechanism of neuropathic pain is extremely complex. Despite of great research efforts on the pathogenesis of neuropathic pain and development of new drugs in recent years, the mechanism of neuropathic pain remains unclear. CCL5 belongs to the C-C chemokine subfamily, and its abnormal regulation of inflammatory responses under pathological condition may induce or exacerbate a variety of diseases. Recently, many researches have suggested that CCL5 has the potential of mediating neuropathic pain, but with unknown mechanism. This paper reviewed the role of CCL5 in the development and regulation of neuropathic pain and discussed the possibility of CCL5 as an cause of neuropathic pain.

Abstract:c-Met is a receptor tyrosine kinase (RTK) with hepatocyte growth factor (HGF) as its natural ligand. c-Met may be activated in a ligand-dependent manner by binding HGF or in a ligand-independent manner. Diverse activation mechanisms of c-Met exist in many malignant tumors, including lung cancer. Moreover, the abnormal activation of c-Met has a close relationship with the occurrence, development and malignant biological behavior of lung cancer. This paper focused on the different activation mechanisms of c-Met in non-small cell lung cancer and small cell lung cancer and also reviewed the research progress of c-Met targeted drugs in lung cancer.

Abstract:Objective To explore the influence of melatonin receptor agonist Neu-P11 on the expression of IRS-1 and GLUT-4 in insulin-resistant 3T3-L1 adipocytes. Methods Insulin resistant 3T3-L1 adipocytes were induced with high glucose/high insulin for 24 hours, and then they were divided into 4 treatment groups: melatonin, Neu-P11, melatonin+luzindole and Neu-P11+luzindole. And non-treated insulin-resistant 3T3-L1 adipocytes were taken as control. Glucose consumption was detected by enzymatic method. IRS-1 and GLUT-4 protein expressions were detected by Western blotting analysis. Results After the insulin-resistant adipocytes were treated with melatonin and Neu-P11, the glucose consumption and the expressions of IRS-1, GLUT-4 proteins were significantly increased compared with the non-treated control group (P<0.05). The expressions of IRS-1, GLUT-4 proteins in melatonin+luzindole(melatonin receptor antagonist) and Neu-P11+luzindole groups were significantly decreased compared with melatonin alone or Neu-P11 alone groups (P<0.05), and were similar to those in the non-treated control group. Conclusion Melatonin receptor agonist Neu-P11 can increase glucose consumption, insulin sensitivity in insulin resistant-adipocytes, which might be associated with the up-regulation of IRS-1 and GLUT-4 protein expression.

Abstract: Objective To observe the effect of plant alkaloid oxymatrine (OMT) on the proliferation and apoptosis of bladder cancer cell line T24 and the possible molecular mechanism. Methods The effects of OMT (0.625, 1.25, 2.5 mg/mL) on the viability and apoptosis of T24 cells were examined by optic, electron microscopy and flow cytometric analysis (FCM). The expression of survivin and caspase-3 genes was detected by RT-PCR, and the protein levels of survivin and caspase-3 were analyzed by Western blotting analysis. Results Typical apoptotic morphologies were observed under optic microscope and electron microscope, and the apoptosis peaks were observed by FCM after treatment with OMT. OMT significantly suppressed survivin expression and up-regulated caspase-3 expression (P<0.05). Conclusion OMT can induce apoptosis of human bladder cancer T24 cell line by decreasing survivin expression and enhancing caspase-3 expression.

Abstract:Objective To explore the clinical characteristics and coronary angiography findings of Tibetan patients with coronary heart disease living in Tibetan plateau area. Methods A total of 117 cases who underwent coronary angiography and were finally diagnosed as coronary heart disease in our hospital were included in this study from September 2006 to October 2011, and they were divided into Tibetan group (76 cases, 65%) and Han group (41 cases, 35%). The clinical data including blood pressure, glucose, blood lipids, blood uric acid and coronary angiography findings were collected and compared. The characteristics of clinical data and coronary angiography of Tibetan group were analyzed. Results There were no significant differences in body mass index, fasting glucose, triglycerides, total cholesterol, low-density lipoprotein cholesterol levels，hypertension, or smoking between the two groups (P>0.05). The onset age, high-density lipoprotein cholesterol and uric acid levels in the Tibetan group was significantly higher than those in the Han group (P<0.05). Single-vessel disease was the dominant type in the Tibetan group, but it also had a larger proportion of complex lesions (three-vessel disease, chronic occlusive lesions, and lesions C). The coronary lesion characteristics were not significantly different between the two groups (P>0.05). Conclusion Hyperuricemia is of great significance for the Tibetan patients with coronary heart disease living in Tibetan plateau area, and the coronary lesions are complex and severe among them.

Abstract:Objective To investigate the skin stimulation effect of essential oils from Ligusticum chuanxiong Hort. Methods HaCaT cells were cultured in vitro. Using MTT and ELISA methods, we examined the effect of the essential oil (different concentrations) on HaCaT cell proliferation and prostaglandin E₂ (PGE₂) levels in culture supernatants of HaCaT cells, and the results were compared with those of oleic acid, a classic permeation enhancer. The cumulative skin stimulation effect was determined by visual scoring in guinea pigs and the histological changes were determined by light microscopy.Results The HaCaT cell viabilities of the 0.005%, 0.015%, and 0.025% essential oil groups were 1.79-, 1.65-, and 1.48-fold that of the 0.005% oleic acid group, respectively, and there was no significant difference between the 0.05% essential oil group and 0.005% oleic acid group. The 0.005%, 0.015%, 0.025%, and 0.05% essential oil influenced the supernatant PGE₂ levels by (0.99±0.08)%,(1.63±0.09)%,(0.98±0.09)%, and (0.04±0.01)%, respectively, which were all significantly lower than the influence of 0.005% oleic acid (\[4.23±0.68\]%, P<0.05). Only slight erythema was observed after continuous administration the essential oil (different concentrations) for 7 days, with no edema or skin uplift, and the erythema caused by 15% essential oil was lower than that caused by 5% oleic acid.Only 15% essential oil exhibited the mechanical injury of the stratum corneum. And 5% oleic acid group showed stripped and lost stratum corneum over large areas. Conclusion Chuanxiong oil is less cytotoxic and less stimulating to the skin compared with oleic acid, and may become an excellent skin permeation enhancer.

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