OBJECTIVETo investigate of the regulatory effect of Rho-kinase pathway activation on angiotensin II (Ang II)-induced contraction of human airway smooth muscle cells (HASMCs) in vitro.

METHODSCultured primary HASMCs were divided into control group, AngII group, AngII + irbesartan group and AngII + Y-27632 group with corresponding treatment. AngII-induced contraction of HASMCs was evaluated using collagen gel lattices and observed morphologically using immunofluorescence assay. Western Blotting was significantly performed to examine the protein expression of Rho-kinase signal pathway.

RESULTSAngII-induced HASMC contraction was inhibited by treatments with irbesartan and Y-27632 as shown by gel contraction assay (P<0.001). Y-27632 treatment produced a stronger inhibitory effect than irbesartan on the expression of phosphorylated moesin, a substrate of Rho kinase (P<0.05).

CONCLUSIONAngII induces the contraction of HASMCs partially as a result of activation of Rho-kinase pathway.

Amides ; pharmacology ; Angiotensin II ; pharmacology ; Asthma ; physiopathology ; Biphenyl Compounds ; pharmacology ; Bronchi ; cytology ; Humans ; Muscle Contraction ; drug effects ; Muscle, Smooth ; cytology ; Primary Cell Culture ; Pyridines ; pharmacology ; Signal Transduction ; drug effects ; Tetrazoles ; pharmacology ; rho-Associated Kinases ; metabolism

OBJECTIVETo analyze the prevalence, clinical features, diagnosis, potential risk factors, anti-tuberculosis treatment efficacy and prognosis of the patients with leukemia complicated with active tuberculosis (TB).

METHODSA retrospective study was performed to analyze the clinical characteristics, relevant examination data, diagnosis methods and follow-up data about 44 leukemia cases complicated with active TB from January 2006 to December 2011 in our single center.

RESULTSThe prevalence of leukemia complicated with active TB was 1.70% (pulmonary TB 1.35%, extra-pulmonary TB 0.35%) and no statistically significant difference was found between each subgroup of acute and chronic leukemia groups (P>0.05). Most of the patients were men, with a male to female ratio of 2.14:1, the median age of 40 years old (range 16 to 78), presenting as atypical clinical manifestations, such as high fever, cough, and so on. Eighteen patients (40.9%) were diagnosed with definitely etiological evidence while the other 26 patients (59.1%) were diagnosed clinically. The extra-pulmonary TB group had a higher purified protein derivative (PPD) test positive rate than that of the pulmonary TB group (88.9% vs 42.9%, P=0.020). The chest CT and T-cell spot of tuberculosis test (T-SPOT.TB) were helpful tools for diagnosis. The potential risk factors included age, sex, nutritional status, neutropenia, decreased cellular immunity, type and course of leukemia, etc. The significant differences in age, gender, administration route of immunosuppressive drugs were found between neutropenic and non-neutropenic groups (P<0.05). The efficacy of first-line anti-tuberculosis therapy was 83.7% and the total course to cure TB was around 12 months. Four patients were dead due to pulmonary TB with a 9.1% attributable mortality.

CONCLUSIONThe prevalence of leukemia complicated with active TB is higher than the general population in our single center. The main characteristics including various potential risk factors, atypical clinical features, diagnoses mainly made by clinical features were found in our patients with leukemia complicated with active TB. However, it showed that these patients demonstrated good responses to the first-line anti-tuberculosis therapy and relative lower attributable mortality.

Adolescent ; Adult ; Aged ; Female ; Humans ; Leukemia ; complications ; epidemiology ; Male ; Middle Aged ; Prevalence ; Retrospective Studies ; Risk Factors ; Tuberculosis ; drug therapy ; epidemiology ; Young Adult

OBJECTIVETo study the clinical manifestations, radiologic findings, pathologic diagnosis and differential diagnosis of primary osteosarcoma in elderly patients.

METHODSTwelve cases of primary osteosarcoma occurring in patients older than 60 years were encountered during the period from 1985 to 2010. The clinical manifestations, radiologic features and pathologic findings were studied and the follow-up data were analyzed.

RESULTSThe sites of involvement included long bones (number = 7), ilium (number = 1), craniofacial bones (number = 2) and soft tissue (number = 2). Radiologic examination showed a mixture of osteosclerotic and osteolytic lesions in 10 patients, soft tissue lesions with high-density areas in 2 patients and soft tissue lesions with periosteal reaction in 8 patients. Histologically, most cases showed features of conventional osteosarcoma. There were 2 cases of malignant fibrous histiocytoma-like osteosarcoma, 2 cases of chondroblastic osteosarcoma and 1 case of well-differentiated intraosseous osteosarcoma. Immunohistochemical study played little role in pathologic diagnosis. Ten patients had undergone amputation, including one patient who had received adjuvant chemotherapy beforehand. Nine patients had follow-up information available. Three of them died of lung metastasis and 1 died of cardiovascular disease.

CONCLUSIONSPrimary osteosarcoma rarely occurs in elderly patients and can easily be missed. Correlation with clinical, radiologic and histologic features is important for arriving at a correct diagnosis.

12E7 Antigen ; Aged ; Antigens, CD ; metabolism ; Bone Neoplasms ; diagnostic imaging ; metabolism ; pathology ; surgery ; Cell Adhesion Molecules ; metabolism ; Chondrosarcoma ; pathology ; Diagnosis, Differential ; Female ; Femoral Neoplasms ; diagnostic imaging ; metabolism ; pathology ; surgery ; Follow-Up Studies ; Humans ; Ilium ; Lung Neoplasms ; secondary ; Lymphoma ; pathology ; Male ; Middle Aged ; Osteitis Deformans ; pathology ; Osteosarcoma ; diagnostic imaging ; metabolism ; pathology ; surgery ; Radiography ; Soft Tissue Neoplasms ; diagnostic imaging ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

Objective To investigate the risk factors, prophylaxis and treatment of de novo malignancies following liver transplantation (LT). Methods Clinical data of 9 patients with de novo malignancies out of 416 patients who underwent LT and received complete follow-up in Liver Transplantation Center, the Third Affiliated Hospital of Sun Yat-sen University from October 2003 to December 2008 were analyzed retrospectively. Seven patients were male and 2 were female. The age ranged from 30 to 60 years old with a median of 57 years old. Four cases had smoking history before operation, 3 cases had long-term passive smoke exposure, and 1 case had family history of hepatocellular carcinoma. The informed consents of all patients were obtained and the ethical committee approval was received. Allogeneic modified piggyback LT was performed in all the patients. The immunosuppressive regimen was methylprednisolone + tacrolimus (FK506) or cyclosporine A (CsA). The patients were followed up after operation and the incidence, treatments and prognosis of malignancies were observed. Results The incidence of malignancies following LT was 2.2% (9/416) including digestive system malignancies (n=3), respiratory system malignancies (n=3), hematological malignancies (n=2) and soft tissue sarcoma (n=1). The elapsed time from LT to diagnosis was 10 to 73 months (median:49 months). The patients received surgery, radiotherapy, chemotherapy accordingly. After the operation, the immunosuppressant of 5 patients changed to sirolimus and 4 patients continued to take FK506 half the dose before tumor diagnosis. Six cases died of tumor progress and multiple organ failure during the follow-up. The interval time from diagnosis of tumor to death ranged from 2 to 25 months (median: 9 months). Conclusions Smoking and use of immunosuppressant may be high risk factors of malignancy following liver transplantation. Normative follow-up and prophylaxis and treatment at the earlier stage are the keys to improve the therapeutic effect.

Objective:To evaluate the effects of different density rat fibroblasts on the expression of conjunctin 43 (Cx43) in cardiomyocytes and cell viability.Methods:Cardiomyocytes and fibroblasts were co-cultured using Transwell, cardiomyocytes were inoculated into the lower chamber of Transwell and fibroblasts into the upper chamber of Transwell.The cells were divided into 3 groups ( n=12 each) by a random number table method: fibroblast density 0.5×10 5 cells/ml group (group C 0.5), fibroblast density 1×10 5 cells/ml group (group C 1), and fibroblast density 2×10 5 cells/ml group (group C 2), with the density of cardiomyocytes 1×10 5 cells/ml in three groups.Cardiomyocytes and fibroblasts were co-cultured for 20 h in three groups.Cardiomyocytes were collected after co-culture for determination of cell viability (by CCK8 method), apoptosis rate (by flow cytometry), and expression of Cx43 mRNA (by quantitative real-time polymerase chain reaction) and expression of Cx43 and phosphorylated Cx43 (p-Cx43) (by Western blot). Results:There was no significant difference in the apoptosis rate of cardiomyocytes among the three groups ( P>0.05). Compared with group C 0.5, the expression of Cx43 protein and mRNA and p-Cx43 was significantly up-regulated in group C 1, the cardiomyocyte viability was significantly increased, and the expression of Cx43 protein and mRNA and p-Cx43 was up-regulated in group C 2 ( P<0.05). Compared with group C 1, the cardiomyocyte viability was significantly increased, and the expression of Cx43 protein and mRNA and p-Cx43 was up-regulated in group C 2 ( P<0.05). Conclusion:Rat fibroblasts up-regulate the expression of Cx43 and enhance the activity of Cx43 in cardiomyocytes and enhance cell viability in a density-dependent manner in a certain range.

Objective:To evaluate the effects of different densities of rat cardiac fibroblasts (RCF) subjected to hypothermic hypoxia-reoxygenation on cardiomyocyte injury and intercellular coupling.Methods:RCF was cultured in vitro and divided into 3 groups ( n=12 each) using a random number table method: RCF density 0.5×10 5 cells/ml group (T 0.5 group), RCF density 1.0×10 5 cells/ml group (T 1.0 group), and RCF density 2.0×10 5 cells/ml group (T 2.0 group). The three groups were placed in an anoxic device, into which 95% N 2 + 5% CO 2 was continuously blown at the speed of 5 L/min for 15 min, and then placed in a 4 ℃ refrigerator for 1 h for low temperature treatment.After completion of culture, cells were placed in a incubator containing 95% air + 5% CO 2 at 37 ℃ for 4 h of reoxygenation.After the end of culture, RCF in three groups were indirectly co-cultured with cardiomyocytes of the same density (1.0×10 5 cells/ml) in a Transwell chamber for 16 h, cardiomyocytes were seeded in the lower chamber of Transwell, and RCF were seeded in the upper chamber of Transwell.After the end of co-culture, cardiomyocytes were collected for determination of the cell viability (by CCK8 method), apoptosis rate (by flow cytometry), expression of connexin 43 (Cx43) mRNA (by real-time fluorescence quantitative polymerase chain reaction), and expression of Cx43 and phosphorylated Cx43 (p-Cx43) (by Western blot). Results:Compared with T 0.5 group, the cell viability, apoptosis rate and expression of Cx43, p-Cx43 and Cx43 mRNA were significantly decreased in T 1.0 and T 2.0 groups ( P<0.01). Compared with T 1.0 group, the cell viability, apoptosis rate and expression of Cx43 and p-Cx43 were significantly decreased ( P<0.01), and no significant change was found in expression of Cx43 mRNA in cardiomyocytes in T 2.0 group ( P>0.05). Conclusions:RCF subjected to hypothermic hypoxia-reoxygenation induces cardiomyocyte injury in a density-dependent manner in a certain range, and the mechanism may be related to down-regulation of the expression of Cx43 and reduction of the activity of Cx43.

Element doping is one of the strategies for increasing the activity of enzyme-like materials.In this study,cobalt,nitrogen and chlorine-doped carbon dots(Co,N,Cl-CDs)were prepared by one-step hydrothermal synthesis utilizing CoCl2 and glutamate as raw ingredients.In weakly acidic environments,Co,N,Cl-CDs exhibited high peroxidase-like activity and could catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine(TMB)by hydrogen peroxide into oxided TMB(oxTMB),resulting in a distinct color change.Free radical scavenging and electron spin resonance techniques were used to further investigate the catalytic reaction mechanism of Co,N,Cl-CDs,and the reactive intermediates produced during the reaction were identified as hydroxyl radical.Acid phosphatase(ACP)could catalyze the hydrolysis of sodium ascorbyl phosphate to release ascorbic acid,which could reduce the blue oxTMB into colorless TMB.Based on this principle,a colorimetric-switching sensing system for detection of ACP was developed by coupling Co,N,Cl-CDs and ACP.The sensing system demonstrated good linearity in ACP activity range of 0.5-20 U/L,with a detection limit of 0.17 U/L.The developed sensing system was used to detection of ACP in real serum samples with good recoveries(97.1%-105.7%)and reproducibility(RSD<5%).The sensing system developed here based on Co,N,Cl-CDs had many advantages such as low detection cost,process visualization,and high sensitivity,providing reference for detection of ACP.

OBJECTIVETo establish a sensitive and effective method for detection of immunoglobulin and T-cell receptor (Ig/TCR) gene rearrangement,and to explore its role in diagnosis and differential diagnosis of lymphoproliferative disorders.

METHODSFifty-eight lymphoid tissue samples from 54 patients with lymphoproliferations were evaluated by the novel BIOMED-2 multiplex polymerase chain reaction (PCR) for antigen receptor genes rearrangement.

RESULTSMultiplex PCR demonstrated monoclonal Ig/TCR gene rearrangements in 22 of 25 (88.0%) B-cell malignancies and 8 of 15 (53.3%) T-cell malignancies. Among 17 benign lymphoproliferations confirmed histopathologically, polyclonal rearrangements were detected in 14 cases (82.4%). In total, the clonality analysis and the final clinico-histopathological diagnosis were concordant in 77.2%. Combination detection of Iglambda and TCR delta gene rearrangements did not increase the detection rate of monoclonal rearrangement of Ig/TCR, but might help to the detection of Iglambda+ or TCR delta+ lymphomas.

CONCLUSIONThe novel BIOMED-2 multiplex PCR strategy is a rapid, reliable and sensitive approach to detecting clonality in suspected lymphoproliferations, especially in atypical cases.

Female ; Gene Rearrangement, B-Lymphocyte, Light Chain ; Gene Rearrangement, T-Lymphocyte ; Humans ; Lymphoproliferative Disorders ; diagnosis ; genetics ; Male ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Objective In the legal proceedings of murder cases,we often encounter the problem of proof standard in traditional forensic pathology in deep research,which is lack of systemic discussion in domestic academic field.This paper aimed to discuss this problem.Methods We compared the relevant characteristics of different legal systems on the standard of proof system,analyzed the elements of the forensic pathology evidence related to murder cases,and discussed the problem with the author's judicial practice experience.Results We believe that the cause of death,death time,injury and injury tools are the three most important forensic pathology evidences in the murder of the standard system as the "three pillars".Conclusions Division of standards in traditional forensic pathology evidence in the murder is of great significance to raise the sense of evidence of forensic medicine,and if it exists major flaws in the evidence of forensic pathology,the evidence can be corrected or the proceedings can be terminated at the very beginning of the litigation phase,which is of great value to the savings of judicial resources.

Objective To investigate the association of 13 single nucleotide polymorphism(SNP)sites in 6 phalange-bone development related genes[fibroblast growth factor receptor 2(FGFR2),indian hedgehog signaling molecule(IHH),Msh homeobox 1(MSX1),Runx family transcription factor 2(RUNX2),SRY-box transcription factor 9(SOX9),Wnt family member 5A(WNT5A)]with human index-ring finger length ratio(2D∶4D).Methods Digital cameras were used to take frontal photographs of the hands of 731 college students(358 males and 373 females)in Ningxia,and image analysis software was used to mark anatomical points and measure finger lengths of index(2th)and ring(4th);genotyping of 13 SNP sites(rs1047057,rs755793,rs41258305,rs3731881,rs3100776,rs12532,rs3821949,rs45585135,rs3749863,rs1042667,rs12601701,rs1829556,rs3732750)for 6 genes by multiplex PCR;One-Way ANOVA or independent sample t-test indirectly assessed the association between 2D∶4D and 13 SNP sites.Results Both left and right hand 2D∶4D were significantly higher in females than males in Ningxia college students(all P<0.01);no statistically significant differences in genotype and allele frequencies of the 13 SNP sites among different sexes(all P>0.05);among different sexes,male left hand 2D∶4D was significantly associated with the genotype of SOX9 gene rs12601701 site(P<0.05)and right hand 2D∶4D was significantly associated with the genotype of WNT5A gene rs1829556 site(P<0.05);the female right hand 2D∶4D was significantly associated with the MSX1 gene rs12532(P<0.01)and rs3821949(P<0.05)sites genotypes.Conclusion SOX9(rs12601701),WNT5A(rs1829556)and MSX1(rs12532 and rs3821949)gene polymorphisms may be associated with the formation of 2D∶4D in Ningxia population.