Terminal restriction fragment length polymorphism(T-RFLP) is a resent molecular approach that can assess subtle genetic differences between strains as well as provide insight into the structure and function of microbial communities. This method overcomes the confinement of conventional culture-dependent methods and has both high sensitivity and throughput making it ideal for comparative analyses. Though there is still no application in our country, more and more investigators are highlighting it. In this article, the fundamental principle of this technique is introduced. The recent application and the development of this technique are also summed up .

Objective To identify the polyreactivity of a monoclonal natural anti-keratin autoantibody 3B4 and to analyze its possible molecular me chanism.Methods enzyme-linked immunosorbent assay (ELISA)and immunohistoche mistry were applied to test the binding reactivity of 3B4 against different anti gens and tissues.The variable region genes and their amino acid composition wer e sequenced.Results 3B4 could reacted with a range of antigens and tissues,i n addition to keratin and skin.The variable region genes of its light chain and heavy chain showed high homology with germline genes VK1 am4 and VH1 J558.42.H CDR3 region,which mainly composed of short side chain amino acids(from 294 to 324 nucleotides around the heavy chain),was the only motif that differs from ot her highly homologous immunoglobulin genes.Conclusions The monoclonal natural anti-keratin autoantibody 3B4,with its variable region genes highly homologo us to germline genes,is highly polyreactive.The flexibility of HCDR3 may contr ibute to the polyreactivity.

?AIM: To study the association of the single nucleotide polymorphism ( SNP) rs1157699 in the calcitonin receptor-like receptor ( CRLR ) gene with primary angle closure ( PAC) in a Han Chinese population.?METHODS: All samples, involved 232 PAC cases and 306 controls, were obtained from an epidemiologic survey conducted in Funing, Jiangsu Province, China. Genotyping were carried out by TaqMan-MGB probe using the real time quantitative polymerase chain reaction system to study the relationship between SNP of rs1157699 in CRLR gene and PAC.?RESULTS: The prevalence of CRLRrs1157699 genotype was 67.4%, 30.0%, 2.6% for CC, CT, TT in cases, and 71.3%, 27.0%, 1.7% in controls respectively.There was no difference between the two groups in the distribution of genotype and allele frequencies of rs1157699 (P>0.05).?CONCLUSION:Our results do not support a significant role for rs1157699 in CRLR with PAC.

Six kinds of saponins (I, II, VII, PA, H) content of 22 samples of Paris polyphylla var. yunnanensis collected from different regions of Yunnan province were determined by HPLC, data was analyzed by SPSS 17. The results showed that the effect of altitude on saponin content was not significant, and the effect of growth area of saponins in P. polyphylla var. yunnanensis was significant, saponin content in sample from west Yunnan was significantly higher than that of samples from other regions.
Altitude ; China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Liliaceae ; chemistry ; Saponins ; analysis

Objective To determine the clinical features,neurophysiological characteristics and cervical magnetic resonance imaging of sensory neumnopathy,and to describe the pathology of skin nerve, sural nerve and spinal dorsal columns.Methods Two patients who died from sensory neuron disease (SND)after infection of digestive tract were discussed including clinical features and ancillary tests which included neurophysiology and pathology of peripheral nerve and spinal dorsal columns.Associated documents are reviewed.Results Early ataxia,widespread sensory symptoms and global loss of deep tendon reflex were the distinctive signs of SND,which was characterized by non-length-dependent abnormalities of sensory nerve action potentials,a hallmark of ganglionopathies.The second patient showed normal cervical magnetic resonance imaging possibly because of short course of disease,while diffuse hyperintensity in the spinal posterior columns of SND was reported.Demyelination of spinal posterior columns and loss of mostly large diameter nerve fibers without regeneration clusters were the main pathological features.Conclusions The distinctive clinical features and neurophysiological characteristics of SND indicate that peripheral sensory nerve fibers are widely damaged.Pathology of spinal posterior columns confirm that central sensory pathway are impaired which allow the localization of the pathologic site to the dorsal root ganglion neurons.Cervical spinal MRI of SND are possibly normal at early phase.

To detect levels of platelet glycoprotein-specific autoantibody in idiopathic thrombocytopenic purpura (ITP), chronic aplastic anemia (CAA), hematologic malignancies and healthy volunteers, and evaluate the clinical significance of platelet glycoprotein-specific autoantibody level in diagnosis for ITP, anti-GPIb/IX, anti-GPIIb/IIIa, anti-GPIV and anti-GPV auto-antibodies were detected contemporaneously by a modified monoclonal antibody immobilization of platelet antigen assay (modified MAIPA). The results showed that the total positive rate of antibodies against platelet GPIb/IX, GPIIb/IIIa, GPIV, GPV were 69.99%, 10%, 20% and 0% in ITP, CAA, hematologic malignancy group and healthy volunteers respectively. There was significant difference between ITP and CAA (chi(2) = 20.71, P < 0.005), between ITP and hematologic malignancy group (chi(2) = 12.22, P < 0. 005). There was no positive finding in the healthy control. It is concluded that platelet glycoprotein-specific autoantibody has high value for the diagnosis of ITP,many kinds of antibodies detection at one time can enhance sensitivity, MAIPA is a specific assay for the diagnosis of idiopathic thrombocytopenic purpura.
Adolescent ; Adult ; Aged ; Autoantibodies ; blood ; Child ; Female ; Humans ; Male ; Middle Aged ; Platelet Membrane Glycoproteins ; immunology ; Purpura, Thrombocytopenic, Idiopathic ; diagnosis ; immunology ; Sensitivity and Specificity

OBJECTIVETo study the hydrodynamic ultrasonic energy extraction technique for Chinese Medicine.

METHODThe content of baicalin in Radix Scutellariae extracts and total flavones in Folium Ginkgo extracts were as indexes respectively and measured by high performance liquid chromatography. We investigated the hydrodynamic ultrasonic energy extraction process for the extraction of Radix Scutellariae and Folium Ginkgo respectively, and compared with refluxing method, decoction and ultrasonic extraction. With the content of the extracts as indexes, Compound prescriptions of Traditional Chinese Medicine were extracted by the hydrodynamic ultrasonic extraction and decoction extraction.

RESULTThe content of baicalin of hydrodynamic ultrasonic energy extracts was significant higher than other extracts. The content of Ginkgo leaves flavones in hydrodynamic ultrasonic energy extracts and ultrasonic extracts had no significant difference. Compared with decocting extracts, hydrodynamic ultrasonic energy extracts of TCM compound prescriptions content was higher greatly.

CONCLUSIONThe hydrodynamic ultrasonic energy extraction process showed higher extraction rate of baicalin of radix scutellariae than traditional extraction methods. And it was beneficial to the dissolution of water-soluble constituents in compound prescriptions of TCM. The content of Ginkgo leaves flavones in the hydrodynamic ultrasonic energy extracts and ultrasonic extracts had no obvious difference.

Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; isolation & purification ; Flavones ; analysis ; chemistry ; Flavonoids ; analysis ; isolation & purification ; Ginkgo biloba ; chemistry ; Reference Standards ; Scutellaria ; chemistry ; Technology, Pharmaceutical ; methods ; Ultrasonics

BACKGROUNDBlocking the 4-1BB/4-1BB ligand (4-1BBL) signal may modulate the secretion of Th1/Th2 cytokines and prolong the survival of the grafts, which play a key role in organ transplantation tolerance. The aim of this study was to investigate the role of blockade of the 4-1BB/4-1BBL co-stimulatory pathway with 4-1BBL monoclonal antibody (mAB) in acute rejection of rat orthotopic liver transplantation.

METHODSThe orthotopic liver transplantation model was set up, while male Lewis rats were used as liver donors and Brown-Norway rats as recipients. The recipient rats were intravenously injected with anti 4-1BBL mAB or isotype control antibody. Groups were monitored for graft survival after transplantation. Plasma chemistry, including aspartate transaminase (AST), alanine aminotransferase (ALT), and bilirubin (BIL), was assayed. The concentrations of interleukin (IL)-2, IL-10 and interferon (IFN)-gamma in plasma were also measured by enzyme-linked immunosorbent assay. Allograft histology images were collected under light microscope and electron microscope.

RESULTSIsotype antibody treated recipients exhibited elevated plasma levels of liver injury markers including AST, ALT and BIL, progressive portal and venous inflammation and cellular infiltration of the liver allografts, and a mean graft survival time (MST) of 10.9 days. Administration of anti 4-1BBL mAB resulted in a decrease in plasma levels of liver injury markers and the concentrations of IL-2, IL-10 and IFN-gamma. The histological grade of rejection on day 7 decreased and MST (17.3 days) increased substantially.

CONCLUSIONSThese results demonstrate that attenuation of acute rejection follows the blockade of the 4-1BB/4-1BBL co-stimulatory pathway with 4-1BBL monoclonal antibody and strongly suggest it is a promising strategy to prevent progression of graft rejection by suppressing T cell-mediated immunity.

4-1BB Ligand ; immunology ; Alanine Transaminase ; metabolism ; Animals ; Antibodies, Monoclonal ; pharmacology ; therapeutic use ; Aspartate Aminotransferases ; metabolism ; Bilirubin ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Graft Rejection ; immunology ; prevention & control ; Graft Survival ; drug effects ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-2 ; blood ; Liver Transplantation ; adverse effects ; Male ; Rats ; Rats, Inbred Lew

OBJECTIVETo investigate the characteristic findings of an anomalous systemic arterial supply to the lung without the pulmonary artery on spiral CT and discuss its nomenclature.

METHODSFour cases of anomalous systemic arterial supply to the left basal segment of the lung without the pulmonary artery were retrospectively reviewed with analysis of the characteristic CT findings.

RESULTSOn spiral CT scans, the involved left lower lung including the entire left basal segments (n=2), the lateral and posterior basal segment (n=1), and the anterior, medial, and posterior basal segment (n=1) had mild volume loss and areas of ground-glass opacity but with normal bronchial trees. The absence of the entire or part of the basal segments of the normal left lower lobar pulmonary artery, anomalous systemic artery originating from the abdominal aorta, diffuse dilatation of the systemic arterial branches distributed in the basal segments of the left lower lobe, and left lower pulmonary venous drainage into left atrium were found in all these patients.

CONCLUSIONThis anomaly presents with characteristic findings on chest spiral CT, for which the nomenclature of local absent pulmonary artery better shows the characteristics of the disease.

Adult ; Aorta, Abdominal ; abnormalities ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Pulmonary Artery ; abnormalities ; diagnostic imaging ; Retrospective Studies ; Terminology as Topic ; Tomography, Spiral Computed

Objective To explore the approach and early effects of endovascular stent-graft deployment in the treatment of portal stenosis of cancerous thrombus.Methods Six cases with portal vein stenosis of cancerous thrombus,which caused by primary hepatic carcinoma(5 cases)and eholangiocarcinoma(1 case)and the severity of stenosis showed on contrast enhanced CT were more than 75% or occluded,were performed percutaneous transhepatie or transsplenic portography.FLUENCY~(TM) endovascular stent-graft(10 mm diameter)was placed at the position of stenosis after gastroesophageal varices embolization.Portal pressure was measured pre-and post-deployment.Results Stents were successfully placed in all patients.The average portal pressure decreased from 50.7 cm H_2O(1 cm H_2O = 0.098 kPa)to 41.3 cm H_2O after endovascular stent-graft deployment.The restenosis were found in 2 cases after one month.Haematemesis and refractory aseites appeared in one case respectively,the other 4 cases showed no significant symptoms above caused by portal hypertension.Conclusion It is safe and feasible for endovaseular stent-graft deployment in the treatment of portal stenosis of cancerous thrombus.Selecting the suitable indications,the symptoms of portal hypertension can be controlled effectively.