The abdominal CT findings of cirrhosis of advanced schistosomiasis in 262 cases were analyzed.All the cases had the calcification in liver in varying degrees,hypodense structure in junction areas and blood vessel in centre,calcification in portal vein system,calcification of colon and so on.The characteristics of CT performance can provide the evidence for defferential diagnosis.

Objective:To construct recombinant lentiviral vectors harboring interference RNA ( RNAi ) targetting murine TNF-αgene,so as to lay the foundation on the RNAi gene therapy.Methods: Three small interfering RNA ( siRNA) sequences targeting murine TNF-αgene ( siRNA1,siRNA2,siRNA3) and negative-control siRNA were designed and synthesized.The inhibition effects of siRNAs on TNF-α,IL-1βand IL-6 secretion of LPS-stimulated RAW264.7 macrophages were observed using real-time PCR and ELISA methods.DNA oligo was designed and synthesized according to the most effective siRNA 2 sequence.The recombinant lentiviral shuttle plasmid expressing short hairpin RNA ( shRNA) was constructed and sequenced.The lentiviral shuttle plasmids with packaging plasmids were transfected into 293T cells to produce lentiviral particles.Results: ①The TNF-αmRNA relative expression levels of siRNA1, siRNA2 and siRNA3 were 0.24±0.01,0.16±0.02,0.19±0.01 respectively,significantly lower than that of negative control (0.95± 0.02) (F=531.3,P<0.001).The inhibition rates at mRNA level were 74.26%,83.09%,79.93%,respectively comparing with negative control.No significance was observed in IL-1βor IL-6 mRNA relative expression change after TNF-αsiRNA transfection ( P>0.05).②The TNF-αprotein expression levels of siRNA1,siRNA2 and siRNA3 were (23.95±1.21),(17.27±1.46),(19.07± 1.57)ng/ml respectively,significantly lower than that of negative control (35.37±2.93)ng/ml (F=18.1,P=0.000 6<0.001).The inhibition rates of protein expression were 32.29%, 51.16%, 46.08%, respectively comparing with negative control.③The PCR product electrophoresis showed that recombinant vectors yielded 343 bp fragments,non-constructed vectors yielded 306 bp fragments.DNA sequencing partially showed insertion sequence.④Lentiviral particles were obtained by transfecting 293T cells with recombinant lentiviral shuttle plasmids and lentiviral packaging plasmids.Cells grew well during virus production with strong fluorescence expression.The titer of concentrated virus was 2×106 TU/μl.Conclusion:The lentiviral vector harboring RNAi targeting murine TNF-αgene has been successfully constructed.

Objective To investigate the effects of lentiviral-mediated RNA interference (RNAi) targeting tumor necrosis factor-α(TNF)-αgene on the expression of TNF-α, interleukin (IL)-1β, IL-6 of murine macrophages RAW264.7, and the efficiency of RNAi experimental gene therapy for the murine collagen-induced arthritis (CIA). Methods The RAW264.7 macrophages were infected by lentivirus-RNAi particles, then stimulated by Lipopolysaccharides (LPS). The TNF-α, IL-1β, IL-6 expression of RAW264.7 macrophages were measured with real-time polymerase chain reaction (PCR) and enzyme linked immunosorbent assay (ELISA). CIA models were esta-blished in DBA/1 mice using bovine type Ⅱ collagen. The treatment effect of lentivirus-RNAi on CIA were observed through arthritis scores, serum TNF-α measurement and hind paw paraffin section hematoxylin/eosin staining after lentivirus-RNAi particles tail vein injection. Results The TNF-αmRNA relative expression level of lentiviral RNAi group was 0.291 ±0.021, significantly lower than that of negative control group 0.925±0.013 (t=25.4, P<0.01). The inhibition rate in mRNA levels was 68.5%. The serum TNF-α level of lentiviral RNAi group was [(249 ±11) ng/ml], significantly lower than that of negative control [(382±6) ng/ml] (t=10.31, P<0.05). The inhibition rate of protein levels was 34.7%. It had no effect on the IL-1β and IL-6 mRNA expression. On the 8th day after systemic administration, the arthritis score of lentivirus-RNAi group was 2.50±0.19, which was significantly lower than that of blank controls (3.63 ±0.18) and negative controls (3.75 ±0.16) (F=42.8, P<0.01). From now on, arthritis score of lentivirus-RNAi group and positive control decreased slowly to at least 2 weeks after treatment induction. The serum TNF-α levels of lentivirus-RNAi group and positive controls were [(35±6) pg/ml] and [(32±7) pg/ml] significantly lower than that of negative controls [(47±3) pg/ml] (t=3.03, 4.11, P<0.01) respectively. Morphological examination showed that the lentivirus-RNAi decreased CIA pathological manifestations. Conclusion Lentiviral-mediated RNAi targeting murine TNF-α gene can effectively inhibit TNF-α expression both in vitro and in vivo, which also effectively improve the CIA arthritis score. Lentiviral-mediated RNAi targeting TNF-αgene provides a potential strategy for rheumatoid arthritis (RA) treatment.

Objective To construct an efficient eukaryotic expression recombinant vector of human interleukin-1O(hIL-lO),and observe its expression in rabbit synoviocytes(RSCs).Methods Total RNA was extracted from peripheral blood mononuclear cells(PBMCs)of a patient with drug allergy.Specific Drimers for full-length open reading frames(ORFs)of hIL-10 were designed according to GeneBank(NM 000572).Withtotal RNA as the template,full-length ORFs of hIL-10 were amplified by reverse transcription Dolymerase chain reaction(RT-PCR).RT-PCR products were digested by restrictive endonucleotidase.then inserted into plasmid pcDNA4/HisMaxA.Both restrictive endonucleotidase analysis and DNA sequencing Were carried out for inserts verification.RSCs were transfected with recombinant plasmid expression vector PcDNA4 HisMaxA-hiL10 by liposome-mediated gene transfer methods,then cultured in vitro.The supernatants were collected af-ter transfection for 12 hours,24 hours,48 hours,72 hours,7 days,14 days respectively for IL-10 measure-ment by enzyme linked immunosorbent assay(ELISA).Results Full-length ORFs of hIL-1o(0.54 kb)had been successfully cloned from PBMCs through RT-PCR.The inserts and insert location of pcDNA4 HisMaxA were in a fight way verified by enzyme analysis and DNA sequencing.ELISA results showed that exogenous hIL-10 gene had expressed in the transfected RSCs from 12 hours to 7 days after transfection,and hIL-10level of transfection group significantly higher than that of the control group.Conclusion pcDNA4 HisMaxA-hiL10,the hIL-10 efficient eukaryotic expression vectors,has been suecessfully constructed.

Objective:To observe the effect of pecking moxibustion on the pattern characteristics of redness,swelling,heat,and pain in the affected joints,also the synovial cell ultrastructure in rats with rheumatoid arthritis(RA)due to damp heat affecting bones/joints,and to explore the anti-inflammatory mechanism of pecking moxibustion in treating the early stage of RA. Methods:Eighteen rats were randomly selected from 78 female ones as the blank group,and all the other rats were subjected to preparing the"differentiation of disease and pattern"RA model due to damp heat affecting bones/joints by using the method of"collagen-induced arthritis plus windy,damp,and hot environment stimulation".Fifty-four rats with successful modeling were randomly divided into a model group,a drug group,and a pecking moxibustion group,with 18 rats in each group.Rats in the drug group were given methotrexate at a dose of 1 mg/(kg·bw)on the 1st,8th,and 15th days.Rats in the pecking moxibustion group were treated with pecking moxibustion at Quchi(LI11),Dazhui(GV14),and Ashi points,and each point was treated with moxibustion for 15 min every day and a total of 3 courses of treatment,with 6 d as a course of treatment.After treatment,the capillary permeability,joint swelling,joint surface temperature,and plantar thermal pain threshold of the diseased joints in rats were observed,and the ultrastructural changes of synovial cells were observed by transmission electron microscopy. Results:The local swelling,surface temperature,and Evans blue(EB)leakage volume were significantly higher(P<0.05),the thermal pain threshold was significantly lower(P<0.05),and the synovial cell ultrastructure was obviously damaged in the affected joints in the model group compared with the blank group.The swelling degree,surface temperature,and EB leakage volume were significantly reduced(P<0.05),the thermal pain threshold was significantly increased(P<0.05),and the ultrastructural abnormalities of synovial cells were significantly improved in the diseased joints in the drug group and the pecking moxibustion group compared with the model group.The thermal pain threshold of rats in the pecking moxibustion group was significantly improved compared with the drug group(P<0.05). Conclusion:Pecking moxibustion obviously improves the pattern characteristics of local redness,swelling,heat,and pain in the diseased joints of rats with RA due to damp heat affecting bones/joints and effectively repairs the ultrastructure of the damaged synovium.It suggests that the pecking moxibustion intervention has a significant anti-inflammatory effect on early RA.

Objective To compare intramedullary nailing assisted by minimally invasive cerclage with simple intramedullary nailing in the treatment of femoral long oblique subtrochanteric fractures.Methods From April 2010 to September 2015,our department treated 39 patients with femoral long oblique subtrochanteric fracture.Of them,16 were treated by cephalomedullary nailing combined with minimally invasive cerclage (observation group of 11 males and 5 females with an average age of 42.8 ± 13.2 years) and 23 by simple cephalomedullary nailing (control group of 17 males and 6 females with an average age of 46.2 ± 10.1 years).Their operation time,intraoperative blood loss,radiologic results (union time and alignment) and functional results [Visual Analog Scale (VAS) and Harris hip score] were compared between the 2 groups.Results The 39 patients were followed up from 12 to 30 months (average,15 months).For the observation group,the varus angle (2.2°± 1.4°) was significantly smaller than for the control group(4.1°±2.2°),the VAS scores at 1 and 3 months postoperatively (3.43 ± 1.54,1.13 ± 1.20) were significantly lower than for the control group (5.61 ± 1.41,3.34 ± 1.82),and the clinical union ratio at 3 months postoperatively(87.5％,14/16) significantly higher than for the control group (47.8％,11/23) (P ＜ 0.05).There were no significant differences between the 2 groups in terms of operation time,intraoperative blood loss,Harris hip score at one year postoperatively,VAS score at 6 months postoperatively,or clinical union ratio at 6 or 12 months postoperatively(P ＞ 0.05).Conclusions Cephalomedullary nailing is effective for the treatment of femoral long oblique subtrochanteric fractures no matter it is assisted by minimally invasive cerclage or not.However,since minimally invasive cerclage has the advantage of improving reduction and mechanical stability,combination of minimally invasive cerclage and cephalomedullary nailing may be more advantageous in early pain-relieving and functional recovery.

Objective To analyze the clinical characteristics of IgG4-related disease (IgG4-RD),guide the selection of therapeutic drugs,and to explore the significance of potential tumor identification for IgG4-RD.Methods A total of 92 patients diagnosed with IgG4-RD and admitted to this hospital from January 1,2017 to December 31,2021were selected as the research subjects by using the Yidu Cloud system.The clinical data conducted the summary analysis. The clinical characteristics of IgG4-RD were summarized.Results The mean age of IgG4-RD was definitely diagnosed in the 92 patients was (58.1±11.3)years old,with 65 male ca-ses (70.7％) and 27 female cases (29.3％).The most commonly affected organ tissues were lymph nodes (37 cases,40.2％),pancreas (33 cases,35.9％) and salivary glands (31 cases,33.7％).In the patients woth the 92 patients,28 cases (30.4％) had involvement of a single organ tissue,while 32 cases (34.8％) had involvement of two or more organs.In the 92 patients,89 cases received steroid therapy,and 71 cases received immunosup-pressive therapy,in which 45 cases (63.4％) used cyclophosphamide.The initial treatment effective rate (72.7％ vs. 55.6％) and one-year non-recurrence rate (38.2％ vs. 20.0％) of the steroid combined immuno-suppressive therapy group were better than those of the single steroid group,but the differences were not sta-tistically significant (P>0.05).The proportion of the patients with tumor comorbidity and IgG4 level>40 g/L (18.2％) was significantly higher than that of the non-tumor comorbidity (1.2％),and the difference was statistically significant (P<0.05).However,there was no statistically significant difference in the proportion of patients with tumor comorbidity compared to the non-tumor comorbidity in other IgG4 level groups (P>0.05).Conclusion IgG4-RD is more common in middle-aged and elderly men,lymph nodes,pancreas and sal-ivary glands are commonly involved,and most patients have the double organs and multiple organs involve-ment. The combination use of hormone and immunosuppressant in treatment is recommended .The IgG4 lev-el>40 g/L in the patients with IgG4-RD may has the suggestive significance for complicating tumor.

Objective:To investigate the rates of low disease activity and clinical remission in patients with systemic lupus erythematosus(SLE)in a real-world setting,and to analyze the related factors of low disease activity and clinical remission.Methods:One thousand patients with SLE were enrolled from 11 teaching hospitals.Demographic,clinical and laboratory data,as well as treatment regimes were collec-ted by self-completed questionnaire.The rates of low disease activity and remission were calculated based on the lupus low disease activity state(LLDAS)and definitions of remission in SLE(DORIS).Charac-teristics of patients with LLDAS and DORIS were analyzed.Multivariate Logistic regression analysis was used to evaluate the related factors of LLDAS and DORIS remission.Results:20.7％of patients met the criteria of LLDAS,while 10.4％of patients achieved remission defined by DORIS.Patients who met LLDAS or DORIS remission had significantly higher proportion of patients with high income and longer disease duration,compared with non-remission group.Moreover,the rates of anemia,creatinine eleva-tion,increased erythrocyte sedimentation rate(ESR)and hypoalbuminemia was significantly lower in the LLDAS or DORIS group than in the non-remission group.Patients who received hydroxychloroquine for more than 12 months or immunosuppressant therapy for no less than 6 months earned higher rates of LLDAS and DORIS remission.The results of Logistic regression analysis showed that increased ESR,positive anti-dsDNA antibodies,low level of complement(C3 and C4),proteinuria,low household in-come were negatively related with LLDAS and DORIS remission.However,hydroxychloroquine usage for longer than 12 months were positively related with LLDAS and DORIS remission.Conclusion:LLDAS and DORIS remission of SLE patients remain to be improved.Treatment-to-target strategy and standar-dized application of hydroxychloroquine and immunosuppressants in SLE are recommended.

OBJECTIVE: The aim of this study was to explore the theoretical framework of cells and the forms of osteogenesis in the mechanism by which demineralized dentin matrix (DDM) induces osteogenesis. METHODS: A total of 24 New Zealand rabbits were used in this study. A total of 4 erector spinae bags were created in each animal. A total of 3 erector spinae bags were implanted with DDM by random selection, whereas the remaining one erector spinae bag was not implanted with DDM. The rabbits were sacrificed after 1, 2, 3, 4, 8, 12, 16, and 20 weeks, and the samples were obtained. The samples were examined by hematoxylin-eosin (HE), tartrate-resistant acid phosphatase (TRAP), and immunohistochemical staining to identify the mesenchymal stem cells, osteoblasts, chondrocytes, and osteoclasts. RESULTS: The results of HE staining showed that in the third week, cartilage- and bone-like matrices, as well as the osteoblast-like cells, were observed. The results of immunohistochemical staining showed that the expressions of CD44, alkaline phosphatase (ALP), and collagen Ⅱ were statistically significant 
(P<0.05). CONCLUSIONS DDM has good histocompatibility and osteoinduction. In addition, induced ectopic osteogenesis mode mainly occurs in the endochondral bone.
Animals ; Bone Matrix ; Dentin ; Osteoblasts ; Osteogenesis ; Rabbits ; Tooth Demineralization