Osteoclasts are derived from pluripotent stem cells in bone marrow and spleen.They play a critical role in inflammation-induced bone loss and joint destruction because in the absence of them,bone de- struction does not occur even when inflammation exists.Synovioblasts in an inflamed joint can secrete numerous inflammatory factors,including tumor necrosis factor alpha(TNF-?)and interleukin-1(IL-1)which not only induce inflammatory reactions but also elevate osteoclast formation and function indirectly or directly through promoting RANKL expression.In this wdy the inflammatory reactions are associated with bone loss and destruction. In this article,we focus on the recent progress in study of TNF-?,IL-1 and osteoclast-target therapies in management of osteoclast-mediated inflammatory bone loss.TNF-?promotes differentiation of osteoclast precursor cells in the peripheral blood and spleen,which causes a marked increase in mature osteoclasts in a diseased joint.However, IL-I supports osteoblast survival and regulates the recombination of osteoclast cytoskeleton,which further stimulates bone resorption.Since osteoclast-target therapies may inhibit osteoclast formation and function,they are becoming more and more important for inflammation-induced bone loss and joint destruction.

Objective To investigate the risk factors of standard selectivity biliary cannulation in endoscopic retrograde cholangiopancreatography (ERCP) for patints with choledocholithiasis,and the effect and safety of assistant cannulation methods.Methods The clinical data were retrospectively analyzed in 372 patients,who received ERCP for choledocholithiasis in the Yijishan Hospital of Wannan Medical College from January 2014 to December 2014.The risk factors of standard biliary cannulation were assessed by univariate and multivariate logistic regression analysis.The success rate of assistant biliary cannulation and the incidence of post-ERCP pancreatitis (PEP) of standard biliary cannulation were compared with doubleguide wire technique (DGT) and precut sphincterotomy (PS),which were used when standard biliary cannulation failed.Results The univariate analysis showed that floppy and long duodenal papilla (P=0.000),angulated common bile duct(P=0.013) were related to failure of standard biliary cannulation.Multivariate analysis showed that floppy and long papilla (P=0.000,OR=0.131,95％CI:0.056-0.307),angulated common bile duct (P =0.003,OR =0.378,95 ％ CI:0.197-0.726) were independent risk factors for standard biliary cannulation.The success rate of standard biliary cannulation was 83.3％ (310/372),62 cases undergone DGT or PS after standard biliary cannulation failure,and the total success rate of cannulation was 99.7％ (371/372).There were 29 cases suffered from PEP in 371 successful cases of biliary cannulation,including 18 cases (5.8％,18/310) of standard biliary cannulation,8 cases (16.3％,8/49) of DGT,and 3 cases (25.0％,3/12) of PS.The incidence of PEP by DGT (x2 =5.532,P =0.019) and PS (x2 =6.994,P=0.008) was significantly higher than that of standard biliary cannulation,but there was no statistical difference on the incidence of PEP between DGT and PS (x2 =0.079,P =0.778).Conclusion Floppy and long duodenal papilla and angulated common bile duct could easily induce the failure of standard biliary cannulation in ERCP for patients with choledocholithiasis.DGT and PS are effective assistant cannulation methods when the standard cannulation fails.

The study is aimed to ensure the quality and safety of medicinal plants by using ITS2 DNA barcode technology to identify Corydalis boweri, Meconopsis horridula and their close related species. The DNA of 13 herb samples including C. boweri and M. horridula from Lhasa of Tibet was extracted, ITS PCR were amplified and sequenced. Both assembled and web downloaded 71 ITS2 sequences were removed of 5. 8S and 28S. Multiple sequence alignment was completed and the intraspecific and interspecific genetic distances were calculated by MEGA 5.0, while the neighbor-joining phylogenetic trees were constructed. We also predicted the ITS2 secondary structure of C. boweri, M. horridula and their close related species. The results showed that ITS2 as DNA barcode was able to identify C. boweri, M. horridula as well as well as their close related species effectively. The established based on ITS2 barcode method provides the regular and safe detection technology for identification of C. boweri, M. horridula and their close related species, adulterants and counterfeits, in order to ensure their quality control, safe medication, reasonable development and utilization.
Base Sequence ; China ; Corydalis ; chemistry ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; chemistry ; genetics ; Molecular Sequence Data ; Nucleic Acid Conformation ; Papaveraceae ; chemistry ; classification ; genetics ; Phylogeny ; Plants, Medicinal ; chemistry ; classification ; genetics

OBJECTIVETo explore the clinical and laboratory features and the prognosis of juvenile dermatomyositis (JDM) complicated with interstitial lung disease (ILD).

METHODData of 39 cases of JDM complicated with ILD hospitalized in Beijing Children's Hospital from January 2005 to December 2011 were collected. The clinical features, laboratory data and prognosis of these children were analyzed.

RESULTOf the 39 cases studied, 16 were boys, and 23 girls. The average age of onset was 5.6 years, and 61.5% of the patients' age of onset (24 cases) was under 6 years. Rashes (17 cases, 43.6%), simultaneous eruption of rashes and muscle weakness (14 cases, 35.9%), fever (4 cases, 10.1%), or muscle weakness (3 cases, 7.7%) were common initial symptoms of the disease. Only 51.3% of the patients (20 cases) had the symptoms of respiratory system, but (24 cases) 61.5% were complicated with that of the gastrointestinal system; (27 cases) 69.2% had at the same time electrocardiographic and echocardiographic abnormalities. The chest high resolution computed tomography (HRCT) showed cord or band-like shadows in their lungs of more than half of the cases (25 cases, 64.1%), and other changes included ground glass-like shadow (10 cases, 25.6%), net and lineation-like shadow (9 cases, 23.1%), nodular change (5 cases, 12.8%). The patients complicated with lung essential infiltration accounted for as high as 71.8% (28 cases). These imaging changes were largely seen on both dorsal sides of their lungs. Severe patients also had mediastinal emphysema, pneumothorax, pneumorrhagia or aerodermectasia. Twenty-four patients underwent pulmonary function examination, and 62.5% of the patients' pulmonary function (15 cases) was abnormal. The fatality rate of the cases studied was 10.1%.

CONCLUSIONThe imaging changes of patients suffering from JDM with ILD were often more severe as compared to the clinical symptoms, and were often complicated with damages to other systems and organs. The prognosis of those patients was poorer than others. Patients with JDM especially at a younger age of onset and with various organ damages should be examined with chest HRCT examinations as early as possible.

Child ; Child, Preschool ; Dermatomyositis ; complications ; diagnosis ; drug therapy ; Female ; Glucocorticoids ; administration & dosage ; therapeutic use ; Humans ; Immunosuppressive Agents ; administration & dosage ; therapeutic use ; Lung ; diagnostic imaging ; pathology ; Lung Diseases, Interstitial ; diagnosis ; drug therapy ; etiology ; Male ; Methotrexate ; administration & dosage ; therapeutic use ; Muscle Weakness ; diagnosis ; epidemiology ; etiology ; Prognosis ; Respiratory Function Tests ; Retrospective Studies ; Tomography, X-Ray Computed

OBJECTIVETo detect the methylation status of p16 gene promotor in DNA derived from plasma and blood cells of patients with systemic lupus erythematosus (SLE) , and it's relationship with clinical symptoms.

METHODSp16 promotor methylation in plasma and peripheral blood cells (PBCs) DNA were simultaneously detected with the methylation specific PCR (MSP) method in 24 active SLE patients, 21 inactive SLE patients, as well as 20 healthy controls.

RESULTSIn the plasma DNA, p16 gene methylation ratio (MP%) was higher in SLE patients than in the healthy controls (64.4% vs. 5.0%, P < 0.05). MP% in the active SLE patients was significantly higher than that in the inactive SLE patients (83.3% vs. 42.9%, P < 0.05). In the PBCs, p16 gene methylation ratio (MC%) in the healthy controls was significantly higher than that in SLE (80.0% vs. 48.9%, P < 0.05). MC% in the active SLE patients (29.2%) was the lowest among three groups. There was no significant difference between the inactive SLE patients and healthy controls (71.4% vs. 80.0%, P > 0.05). Each patient could be judged as one of the four methylation patterns: MP/MC, UP/MC (UP: unmethylated plasma p16) , MP/UC (UC: unmethylated PBCs p16) , and UP/UC. The ratios of MP/ MC and UP/UC were similar between the active and inactive SLE patients. However, different distributions of other two patterns were found in the active and inactive SLE patients as UP/MC 4.2% vs. 42.9% (P <0.05) and MP/UC 58.3% vs. 14.3% (P < 0.05), respectively. The active SLE patients with MP/UC and the inactive SLE patients with UP/MC showed different clinical symptoms and laboratory examinations. Significant correlation was found between the disease activity index for lupus patients (SLEDAI) scores and MP% (r = 0.93), between the SLEDAI scores and MC% (r = - 0.96) also between MC% and MP% (r = - 0.79).

CONCLUSIONThe p16 methylation assay provides available information for the diagnosis, judgment of disease activity, as well as novel insights into the pathogenesis underlying this disease.

Adult ; DNA Methylation ; Female ; Genes, p16 ; Humans ; Lupus Erythematosus, Systemic ; diagnosis ; genetics ; Male ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; genetics

OBJECTIVETo investigate the expressions of chemokine receptors and interleukin (IL) receptors on the peripheral blood mononuclear cells (PBMCs) from systemic lupus erythematosus (SLE) patients and their correlations with clinical features as well as SLE disease activity index (SLEDAI).

METHODSThe mRNA expressions of chemokine receptors and IL receptors on PBMCs of 93 SLE patients and 30 healthy controls were detected by reverse transcription-polymerase chain reaction, including CCR2, CCR3, CCR4, CCR5, CCR6, CCR8, CXCR3, CXCRS, CX3CR1, XCR1, IL-4R, and IL-10R. The clinical features of SLE patients were recorded. The correlations of chemokine receptors and IL receptors mRNA expressions with clinical features as well as SLEDAI were assayed using linear regression analysis.

RESULTSThe level of CCR5 mRNA in SLE patients (including active and inactive SLE) was significantly higher than that in healthy controls (P < 0.05), and there was no significant difference between active and inactive patients in this respect (P > 0.05). CX3CR1 mRNA expression significantly increased from healthy control to inactive SLE to active SLE in sequence. The others (except for CCR8, CXCR3, and IL-10R) in active SLE patients were significantly higher than those in both inactive SLE patients and healthy controls (all P < 0.05). There were positive correlations between SLEDAI and CCR2 (r = 0.424, t = 4.313, P < 0.001), CCR3 (r = 0.518, t = 5.410, P < 0.001), CCR4 (r = 0.376, t = 3.851, P < 0.001), CCR6 (r = 0.457, t = 4.513, P < 0.001), CXCR5 (r = 0.455, t = 4.629, P < 0.001), CX3CR1 (r = 0.445, t = 4.523, P < 0.001), as well as XCR1 (r = 0.540, t = 5.445, P < 0.001). And CCR5 mRNA expression level was positively correlated with IL-4R mRNA (r = 0.313, t = 2.353, P < 0.05). The patients with myositis and cutaneous vasculitis simultaneously showed lower levels of CCR5 and CX3CR1, and CCR5 expression was negatively correlated with the scores of SLEDAI in SLE cases accompanied by photosensitivity (r = 0.426, t = -2.155, P < 0.05).

CONCLUSIONIncreased expressions of CCR5 and CX3CR1 on PBMCs may be indicators in clinical survey for SLE.

Adolescent ; Adult ; CX3C Chemokine Receptor 1 ; Child ; Female ; Humans ; Leukocytes, Mononuclear ; immunology ; Lupus Erythematosus, Systemic ; etiology ; immunology ; Male ; Middle Aged ; RNA, Messenger ; blood ; Receptors, CCR5 ; genetics ; Receptors, Chemokine ; genetics ; Receptors, Interleukin-10 ; genetics ; Receptors, Interleukin-4 ; genetics

Saliva and blood were collected from two patients who had not received post exposure prophylaxis in the cities of Wenzhou and Xinning respectively. Both patients were confirmed as positive for rabies by detection of rabies virus specific nucleoprotein antibodies in the sera by Western Blot. However, rabies virus specific RNA was only identified in the saliva collected from the patient in Wenzhou. Furthermore, the isolate Zhejiang Wz0 (H) was obtained by inoculating one-day-old suckling mice. Both nucleoprotein (N) and glycoprotein (G) genes from the isolate were amplified by RT-PCR and sequenced. Phylogenetic analysis indicated that the isolate belonged to classic rabies virus, and shared a higher homology with the street viruses from dogs in the main endemic areas in China and the street virus from dogs in Indonesia than with other known strains. Further comparison of the deduced amino acid sequences between the isolate and the vaccine strains used in China showed that the virus had a higher level of homology with the vaccine strain CTN than with the other vaccine strains (3aG, PV, PM and ERA). In particular, amino acid residues substitutions located in antigenic site Ⅲ in the G protein, which could react with the neutralizing antibodies, were observed. These results suggested that the virus belonged to the classic rabies virus, and both N and G genes diverged from the current vaccine strains used in China at either the nucleotide or the amino acid level.

Objective To explore the feasibility of robot-assisted radical prostatectomy(RARP)in the treatment of highly suspected prostate cancer without biopsy,in order to provide reference for clinical practice.Methods This study included 82 patients with highly suspected prostate cancer treated in our hospital during Jan.2021 and Dec.2022.The patients were divided into the puncture group(n=41)and non-puncture group(n=41)with random number table method.All patients underwent RARP.Biopsy was performed before operation in the puncture group while no biopsy was performed in the non-puncture group.The main outcome indicators(diagnostic accuracy,complications,hospital stay,hospital costs,urinary control rate)and secondary outcome indicators(operation time,intraoperative blood loss,number of blood transfusion,catheter indwelling time,positive incisal margin,anxiety and depression scores before and after treatment)were compared between the two groups.Results All 41 cases(100％)in the puncture group were prostate cancer,and 40 cases(97.56％)in the no-puncture group were pathologically confirmed as prostate cancer.The non-puncture group had significantly lower perioperative complication rate(19.52％vs.2.50％),shorter operation time[(75.31±20.35)min vs.(60.25±10.64)min],less intraoperative blood loss[(141.25±30.45)mL vs.(111.45±33.58)mL],shorter hospital stay[(7.89±1.21)d vs.(5.13±1.02)d],lower total hospital costs[(4.12±0.56)ten thousand yuan vs.(2.61±0.43)ten thousand yuan],and lower anxiety and depression scores[(7.02±1.25)vs.(1.99±0.42);(7.05±1.31)vs.(1.95±0.38)](P＜0.05).The no-puncture group had slightly higher urinary control rate 1,6 and 12 months after treatment than the puncture group(75.00％vs.58.54％;87.50％vs.80.49％;95.00％vs.92.68％),but with no statistical significance(P＞0.05).Conclusion RARP is feasible in the surgical treatment of prostate cancer without biopsy,which can shorten the postoperative rehabilitation time,reduce the hospital costs,and alleviate the anxiety and depression of patients.

BACKGROUNDRecent studies have suggested that mature T cells can change their specificity through reexpression of recombination-activating genes (RAG) and RAG-mediated V(D)J recombination. This process is named receptor revision and has been observed in mature peripheral T cells from transgenic mice and human donors. However, whether thebreceptor revision in mature T cells is a random or orientated process remains poorly understood. Here we used the Jurkathuman T cell line, which represents a mature stage of T cell development, as a model to investigate the regulation of Tcell receptor (TCR) gene recombination.

METHODSTCR Dbeta-Jbeta signal joint T cell receptor excision DNA circles (sjTRECs) were determined by nested and seminested PCR. Double-strand DNA breaks at recombination signal sequences (RSSs) in the TCRVbeta chain locus were detected by ligation-mediated-PCR. Further analysis of the complementarity-determining region 3 (CDR3) size of the TCRVbeta chain was examined by the TCR GeneScan technique.

RESULTSRAG1, RAG2, and three crucial components of the nonhomologous DNA end-joining (NHEJ) pathway were readily detected in Jurkat. Characteristics of junctional diversity of Dbeta2-Jbeta2 signal joints and ds RSS breaks associated with the Dbeta2 5' and Dbeta 2 3' sites were detected in DNA from Jurkat cells. CDR3 size and the gene sequences of the TCRVbeta chain did not change during cell proliferation.

CONCLUSIONSRAG1 and RAG2 and ongoing TCR gene recombination are coexpressed in Jurkat cells, but the ongoing recombination process may not play a role in modification of the TCR repertoire.However, the results suggest that Jurkat could be used as a model for studying the regulation of RAGs and V(D)J recombination and as a "special" model of the coexistence of TCR gene rearrangements and "negative" receptor revision.

Antigens, Nuclear ; genetics ; Base Sequence ; Complementarity Determining Regions ; DNA Breaks ; DNA-Binding Proteins ; genetics ; Genes, RAG-1 ; Genes, T-Cell Receptor ; Humans ; Jurkat Cells ; Ku Autoantigen ; Leukemia, T-Cell ; genetics ; Molecular Sequence Data ; Nuclear Proteins ; genetics ; Recombination, Genetic

OBJECTIVETo explorer the effectiveness of enriched bone marrow stem cells technique for lumbar fusion.

METHODSWith the randomization and control principles, 2 graft materials [Enrichment bone marrow mesenchymal stem cells hybridized with beta-tri calcium phosphate (composite graft group), autologous iliac crest bone graft (autograft group)] were compared in posterior lumbar fusion procedures. 56 patients with degenerative disc disease, lumbar instability or spinal stenosis, were included. The volume of cells suspension in pre- and post-enrichment and the number of nucleated cells (NCs) were identified. The number of osteoprogenitor cells was estimated by counting the colony-forming units which express alkaline phosphatase (CFUs/ALP+). Then the efficiency of the enrichment was evaluated. Clinical follow-up with roentgenogram and Oswestry scale scores was performed for outcome evaluation.

RESULTS(249 +/- 31) ml bone marrow per patient from bilateral iliac crests was aspirated peri-operatively. About (43 +/- 11) ml enriched bone marrow was collected. The number of NCs was concentrated from (15.9 +/- 3.3) x 10(6)/ml to (44.1 +/- 10.8) x 10(6)/ml, CFUs/ALP+ was significantly increased from (118 +/- 86)/ml to(486 +/- 305)/ml. The follow-up was about (26.3 +/- 7.5) months. There was no significant differences in age, gender, disease and fusion segments between the two groups. The fusion rate was 93.3% and 96.2% for composite graft group and autograft group, respectively (chi2 = 0.2146, P = 0.6432). There was no difference in operation time between the two group (t = 0.5243, P = 0.6022), but blood loss in composite graft group was more than that in autograft group (t = 6.4664, P < 0.01). Cell salvage for auto-transfusion could transfuse back half of the blood loss during operation. No hematoma or chronic soreness in the bone marrow donor sites of composite graft group occurred, but a little exudation or moderate swelling in the wound happened in 4 cases which disappeared under medical treatment. Meanwhile, 15.4% patients had hematoma in the iliac bone donor site and 26.9% patients had chronic soreness, but no case had wound problem in autograft group. As for Oswestry scale scores, there was no significant difference between the two groups.

CONCLUSIONSThe enrichment technique of autologous bone marrow stem cells can greatly increase the concentration of MSCs. It is a rapid and safe method used peri-operatively. The composite material of enriched MSCs and porous beta-TCP is a good bone substitute in posterior spinal fusion.

Adult ; Aged ; Bone Substitutes ; Bone Transplantation ; methods ; Calcium Phosphates ; Female ; Follow-Up Studies ; Humans ; Ilium ; transplantation ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Spinal Fusion ; instrumentation ; methods ; Stem Cell Transplantation ; Transplantation, Autologous ; Treatment Outcome