1.The inhibition of common immunosuppressive drugs on the development of de novo donor specific antibodies
Lei FU ; Zhengsheng RAO ; Turun SONG ; Dongyang ZENG ; Tao LIN
Chinese Journal of Organ Transplantation 2017;38(7):390-396
Objective To explore the inhibition of different immunosuppressive drugs or immunosuppressive combination regiments on the development of de novo donor specific antibodies (dnDSA).Methods We used BABL/c mice and C57BL/6 mice alloimmunized using donor splenocytes for establishing model of producing dnDSA.After that,mice were divided into 10 groups,and were administrated with mycophenolate mofetil (MMF,300 mg· kg-1 · d-1);sirolimus (0.3 mg·kg-1 ·d-1);tacrolimus(0.9 mg· kg-1 · d-1);cyclosporine A (45 mg· kg-1 · d-1);MMF (150 mg· kg-1· d-1) + tacrolimus(0.45 mg · kg-1 · d-1);MMF (150 mg · kg-1 · d-1) + cyclosporine A (22.5 mg·kg-1 ·d-1);MMF (150 mg·kg-1 ·d-1) + sirolimus (0.15 mg·kg-1 ·d-1);sirolimus (0.15 mg· kg-1 · d-1) + tacrolimus (0.45 mg· kg-1 · d-1);sirolimus (0.15 mg· kg-1 · d-1) + cyclosporine A (22.5 mg· kg-1 · d-1);and placebo;respectively.Anti-serum was harvested and tested using flow cytometry.Results (1) Four weeks after sensitization,the dnDSA level in BALB/c and C57BL/6 group was 88.86% and 25.58%.Comparing with control group(25.33%),there was a significant increase in BALB/c group(89.23% versus 25.33%;P<0.001),whereas no change was found in C57BL/6 group (25.58% versus 25.33%;P =0.259),that we chose BABL/c mice as the model for sensitization.(2)After 4 weeks of sensitization and immunosupression,dnDSA level was 29.31%,46.33%,57.10% and 66.35% in MMF,sirolimus,cyclosporine A and tacrolimus monotherapy group respectively.In comparison to sensitization group,dnDSA level in all monotherapy arm reduced significantly.Intra-group analysis found MMF monotherapy had the most effective inhibition of dnDSA production,sirolimus came the second,and tacrolimus and cyelosporine A had equivalent effectiveness.dnDSA level was 31.00%,33.12%,45.18%,44.62% and 61.60% in MMF+ sirolimus,MMF+ tacrolimus,MMF+ cyclosporine A,sirolimus + cyclosporine A and sirolimus + tacrolimus combination treatment arm,respectively,which was much lower than that of sensitization group (P<0.001,P<0.001,P<0.001,P<0.001,P =0.015).MMF + tacrolimus and MMF+ sirolimus combination treatment had comparable effectiveness in dnDSA inhibition,which is greater than that of the rest of three combination.Conclusion Our data show that the effects of single immunosuppressive drugs on the level of dnDSA is MMF>sirolimus>cyclosporine A =tacrolimus,and that in combination regimens is MMF + tacrolimus =MMF + sirolimus>MMF + cyclosporine A =sirolimus + cyclosporine A =sirolimus + tacrolimus.In conclusion,this study demonstrates that MMF+ tacrolimus combination treatment arm can minimize the development of dnDSA.But,this conclusion remains to be confirmed by future clinical studies.
2.Expression,Purification and Bioactivity Identification of IFN-tau
Jia-Lei FU ; Chang-Zheng SONG ; Geng-Lin ZHANG ;
China Biotechnology 2006;0(03):-
Interferon-tau (IFN-tau) is a newly discovered IFN of type I. It was originally found for its role as a pregnancy recognition hormone in ruminant animals such as sheep and cows. Like the other type I IFNs, IFN-tau have the same biological activities including antiviral, antiproliferative and immunomodulatory effects. In order to clearly identify the function of IFN-tau, the coding sequence of IFN-tau was amplified by PCR from IFN-tau cDNA, this fragment digested by EcoR I and BamH I and was inserted into pBV220 to form the recombinant plasmid pBV220/IFN-tau which was then transformed into E.coli BL21. It was found that pBV220/IFN-tau was highly expressed as inclusion body in BL21. Next, the expressed protein was purified on S-100 High Resolution and the purified product was confirmed by amino acid sequence analysis. Moreover, the standard antiviral activity test indicated that the activity of IFN-tau was about 2.09?106IU/ml.
3.Study on Compatibility of TCM Prescriptions for Treating Alzheimer Disease Based on Data ;Mining
Sen LIN ; Jie LU ; Kun SONG ; Zongfu CHEN ; Sen JIANG ; Chunliang PAN ; Xuebo LI ; Xianjun FU
Chinese Journal of Information on Traditional Chinese Medicine 2015;(5):41-44
Objective To study the compatibility of TCM prescriptions of TCM practitioners of all dynasties of Alzheimer disease (AD). Methods Amnesia, forgetting, dementia, and idiot were set as search words to retrieve relevant literature in Encyclopadia of Traditional Chinese Medicine. Prescription information was screened and standardized to build database. Frequency analysis and association rules were used to mine TCM prescriptions and compatibility rules. Results Totally 449 AD related prescriptions were selected, involving 682 Chinese medicinal herbs. The individual herb with the highest frequency was Ginseng Radix Rhizoma (192);the herbal pair with the highest frequency was Ginseng Radix Rhizoma-Polyhalae Radix (182);the herbal combination with 3 Chinese medicinal herbs with the highest frequency was Poria with Hostwood-Ginseng Radix Rhizoma-Polyhalae Radix (79);the herbal combination with 4 Chinese medicinal herbs with the highest frequency was Polyhalae Radix-Ginseng Radix Rhizoma-Poria with Hostwood-Glycyrrhizae Radix et Rhizoma (37). The results of association rules showed that Ginseng Radix Rhizoma-Polyhalae Radix, Ginseng Radix Rhizoma-Glycyrrhizae Radix et Rhizoma, and Ginseng Radix Rhizoma-Poria with Hostwood were commonly used compatibilities in AD related prescriptions. Conclusion Treatment of TCM practitioners in all dynasties for AD mainly chooses Chinese medicinal herbs with the efficacy of tonifying qi and soothing nerves. The compatibilities and combinations are reasonable and with certain representativeness.
4.Expression and Antigenic Characterization of the Epitope-G1 of the Bovine Ephemeral Fever Virus Glycoprotein in Pichia pastoris
Fu-ying, ZHENG ; Guo-zhen, LIN ; Chang-qing, QIU ; Kui-zhang, YUAN ; Jun-ying, SONG
Virologica Sinica 2007;22(5):347-352
The epitope-G1 gene of Bovine ephemeral fever virus (BEFV) glycoprotein was synthesised by PCR and cloned into expression vector pPIC9K to construct recombinant plasmid pPIC9K-G1. Then the pPIC9K-G1 was linearized and transformed into Pichia pastoris GS 115. The recombinant P. pastoris strains were selected by a G418 transformation screen and confirmed by PCR. After being induced with methanol, an expressed protein with 26 kDa molecular weight was obtained, which was much bigger than the predicted size (15.54 kDa). Deglycosylation analysis indicated the recombinant G1 was glycosylated. Western blot and ELISA tests, as well as rabbit immunization and specificity experiments indicated that the target protein had both higher reaction activity and higher immunocompetence and specificity. The recombinant G1 protein could be used as a coating antigen to develop an ELISA kit for bovine ephemeral fever diagnosis.
5.The safety of Habib VesOpen bipolar radiofrequency ablation catheter used in the treatment of portal vein tumor thrombus:an experimental study in miniature pig models
Lin ZHANG ; Jinxin FU ; Peng SONG ; Kai YUAN ; Jieyu YAN ; Feng DUAN ; Maoqiang WANG ; Fengyong LIU
Journal of Interventional Radiology 2015;(6):515-519
Objective To explore the safety of Habib VesOpen bipolar radiofrequency ablation (RFA) catheter used in the treatment of portal vein tumor thrombus (PVTT). Methods A total of 10 miniature pigs were randomly divided into 3 groups. Group A(n=6):RFA of normal portal vein was directly performed;group B (n=2): balloon obstruction of the portal vein was performed first, which was followed by RFA for the fresh thrombus in the portal vein; group C (n=2): PVTT model was established first, and RFA of the portal vein was carried out when the portal thrombus became organized. MRI examination was employed at one, 3 and 4 weeks after RFA; the animals were sacrificed 4 weeks after RFA and pathological examination of portal vein was performed. Results Pigs of group A received portal vein RFA under the condition of 5 W power for 0.6-3.6 min. No obvious abnormality was detected by MRI and pathological examination , which were performed one month after the treatment. In the pigs of group B , MRI performed after RFA showed that the damage of portal vein area was more serious than that in the pigs of group A;abdominal MRI examination performed at one, 3 and 4 weeks after RFA showed that the portal venous edema was gradually decreased;pathological examination at one month after RFA demonstrated serious injury of adjacent liver tissue. Pigs of group C received portal vein RFA under the condition of 7 W power for 1.5 min; no obvious edema of the ablated area was observed on MRI performed after RFA , and pathological examination revealed organized thrombus necrosis and va scular endothelial cell damage. Conclusion When Habib VesOpen bipolar RFA catheter is used for the treatment of PVTT, the RFA power and time should be properly selected according to the severity of PVTT. In order to ensure a safer procedure, high power and short ablation time should be used when the severity of PVTT is mild, while low power and longer ablation time are recommended when the PVTT is more severe.
6.Volumetric changes of upper airway after rapid maxillary expansion:A Meta analysis
Lin SONG ; Weiwei XIAO ; Tengfei FU ; Meng GAO ; Zaixiu QIN ; Yongming LI
Journal of Practical Stomatology 2017;33(3):358-363
Objective:To systematically evaluate the volumetric changes of upper airway after rapid maxillary expansion using Meta analysis.Methods:All literatures about volumetric changes in the upper airway after rapid maxillary expansion were searched from the database in general.The literatures were screened according to the correlation and the inclusion criteria,included the literatures were analyzed by Rev Man 5.3.Results:11 literatures were finally selected.Meta analysis indicated that,after treatment nasopharyngeal volume increased by 0.62 cm3(P=0.000 2),palatopharyngeal volume increased by 0.62 cm3(P=0.02),glossopharyngeal volume increased by 0.39 cm3(P=0.11),oropharyneal volume increased by 0.40 cm3(P=0.27).Conclusion:The existing evidence indicates that rapid maxillary expansion can increase the volume of nasopharynx and retropalatal part of upper airway.
7.Expression of virus-like particles of enterovirus 71 in Hansenula polymorpha
Meirong GU ; Linlin SONG ; Shanshan XU ; Zuoshen FU ; Fuyu LIN ; Xianchen ZHANG ; Wenjin WEI ; Shiru JIA
Chinese Journal of Microbiology and Immunology 2013;(8):604-609
Objective To express virus-like particles(VLP) of enterovirus 71 (EV71) in Han-senula polymorpha.Methods The coding sequences of P1 and 3CD genes of EV71 were optimized accord-ing to codon usage bias of Hansenula polymorpha for achieving high expression , and then cloned into the ex-pression vector PMV of Hansenula polymorpha .The recombinant expression vector PMV-P1-3CD was trans-formed into Hansenula polymorpha AU 0501 .The transformants were stably cultured in selective medium (Yeast Nitrogen Base) and screened for strains with positive P1 and 3CD genes by PCR.Then an induced cultivation on the recombinant strains were performed in a medium supplemented with methanol to a final concentration of 1.0%and the expressed products were analyzed by SDS-PAGE and Western blot assays to select high expression strains .The high expression strains were cultured in 30 L fermentor and its fermenta-tion products were analyzed by electronic microscope after purification .Results EV71 recombinant expres-sion strains were successfully constructed .The results of SDS-PAGE showed that the expressed products had obvious VP3, VP1, VP0 protein bands with molecular weights of 26×103, 33×103 and 35×103, respective-ly, which were consistent with the expected molecular weight of the fusion proteins .Western blot demonstra-ted that the expressed products could be specifically recognized by the polyclonal antibody against EV 71-VP1 at 33 ×10 3 , indicating its high immunoreactivity .ELISA confirmed that the expression level of EV 71 fermen-tation products was reached to 200 mg/L.Electronic microscope analysis showed that the VLP of recombi-nant EV71 were 24-30 nm in diameter with normal structure .Conclusion The virus-like particles of human enterovirus 71 are successfully expressed in Hansenula polymorpha , which provides a foundation for the fur-ther development of EV 71 VLP vaccine .
9.Correlative factors influencing acetabular fracture associated with femoral nerve injury
Yue FANG ; Yan-Cheng SONG ; Tian-Fu YANG ; Lei-Ting CHI ; Guang-Lin WANG ;
Chinese Journal of Trauma 2003;0(08):-
Objective To improve the effect of clinical diagnosis and treatment of acetabular fracture with a femoral nerve injury by analyzing the causes of femoral nerve injury following acetabular fractures.Methods From January 1996 to November 2004,146 cases of acetabular fractures were treated operatively.Six cases of them were complicated with femoral nerve injury.The causes of femoral nerve injury were analyzed on the basis of clinical manifestations,CT scan and 3-dimensional reconstruc- tion.All the cases were classified according to Letournel and Judet classification.Three cases had hema- toma compression (2 cases with double column fractures and 1 with transverse-posterior wall fractures);2 cases had femoral nerve injury caused by fracture fragments (1 case with anterior wall fracture following anterior hip dislocation,the another with old fracture of anterior column combined with fracture of superior ramus of pubis);and one case had anterior column fracture combined with fracture of wing of ilium,and the femoral nerve was injured by traction in operation.Clearance of hematoma,nerve tract decompression and epineuria solution were performed in 5 cases,and 1 case was treated conservatively.Results The average follow-up period was 1.8 years(ranging from 1 to 3 years).The muscle power of quadriceps fem- oris recovered from 1-2 grade before operation to 4-5 grade after operation in 5 cases.The function of ex- tensor knee and gait was normal.The function of sensory completely recovered in 4 cases.One case was followed up for 2 years,which showed the patient still suffered from hypoesthesia in the lower 2/3 of the thigh and the medial of the leg.One ease of traction injury was followed up for 1.2 years,showing the muscle power recovered to normal,but still presented with sensory disability.Conclusion Acetabular fractures associated with femoral nerve injury are rare.For complex acetabular fractures and severe trau- ma,attention should be given to the possibility of femoral nerve injury.Fragment stabbing and compres- sion of hematoma around iliopsoas muscle are the common causes of femoral nerve injury following ace- tabular fractures.Iatrogenic injury should not be ignored.
10.Study of the therapeutic effects of free radical scavenger edaravone on cerebral hemorrhage in rats
Ling-Lin DONG ; Fu-Qiang GUO ; You-Song YANG ; Hong-Yuan DAI ; Wen-Bin WU ;
Chinese Journal of Neurology 2001;0(01):-
Objective To explore therapeutic effects and mechanisms of radical scavenger edaravone on experimental cerebral hemorrhage.Methods Two hundred-forty male SD rats were divided randomly into four groups:control group,cerebral hemorrhage group,edaravone treatment group before operation (A) and edaravone treatment group after operation (B).Experimental cerebral hemorrhage model was made according to the method reported by Rosenberg.Water quantity contained in brain and nervous missing sign were observed,meanwhile the levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in brain tissue were measured.Results Compared with cerebral hemorrhage group,nervous missing sign and water quantity contained in brain obviously changed in edaravone treatment group (P