1.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.
2.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.
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.
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.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 .
7.The diagnosis and treatment of ischemic bowel disease,experience in 73 cases
Wei FU ; Chao-Lai MA ; Zi-Shun ZHANG ; Min LYU ; Xue-Song YANG ; Tong-Lin ZHANG
Chinese Journal of General Surgery 1993;0(02):-
Objective To analyze clinical features and sum up experience for the treatment of ischemic bowel disease. Methods Clinical data of 73 patients with the diagnosis of ischemic bowel disease were retrospectively analyzed. ResultsTwenty-eight patients were male and 45 patients were female. The median of age was 65 years (range of 38 to 89 years). Forty-eight patients were associated with hypertension, 23%(17/73) patients had a history of coronary disease and 15% (11/73) had diabetes. Seventy patients presented symptom of abdominal pain and 93% (68/73) had hematochezia. Symptoms relieved by conservative treatment in 96% (63/66) patients. Nine patients underwent a surgery. One patient died of sepsis postoperatively. One suffered from colostomy necrosis and leakage of the rectum segment. Conclusion 1. Elder patients presenting symptoms of abdominal pain and hematochezia, especially with a history of cardio-cerebrovascular disease and diabetes should be considered for the possibility of ischemic bowel disease. 2. Most patients with ischemic bowel disease could be successfully treated by conservative therapy. 3. Surgery for patients with chronic relapsing and nonresponsible symptoms was difficult and patients often suffer from high postoperative complications.
8.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.
9.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.
10.TCM practitioners’ attitudes and perceptions regarding the use of Ephedra sinica Stapf: An observational study
Aiwen Chang ; Xiaopeng Zhao ; Lin Zhang ; Sijia Zhao ; Zhongyi Pan ; Chenxi Song ; Yanling Fu
Journal of Traditional Chinese Medical Sciences 2024;11(4):435-442
Objective:
To understand the attitudes and perceptions of traditional Chinese medicine (TCM) practitioners in Beijing TCM hospitals regarding the use of Ephedra sinica Stapf (E. sinica, Ma Huang).
Methods:
A two-stage cross-sectional study was conducted using a questionnaire survey of TCM practitioners in Beijing TCM hospitals between April 2023 and March 2024. The questionnaire included demographic information, the clinical background of TCM practitioners, and the clinical application of E. sinica. Logistic regression analysis was used to analyze the relevant influencing factors when using E. sinica.
Results:
Of the 465 questionnaires collected, 441 were valid. Among these, 84.81% (374/441) reported having used E. sinica in clinical practice at least once. The commonly used doses of E. sinica—excluding the pediatric department—were 10 g for high doses, 6 g for medium, and 3 g for low. The three most frequently used formulas for E. sinica included Maxing Shigan decoction, Mahuang decoction, and Xiao Qing Long decoction. The most common TCM patterns treated with a high dose of E. sinica were wind-cold exterior pattern, wind-cold invading the lung, and wind and water combat with meridians congealed by cold. The top three Western medical diagnoses when using E. sinica for treatment were common cold, pneumonia, and upper respiratory tract infections. Nearly half of the respondents reported experiencing adverse reactions from the oral administration of E. sinica, with the most common being palpitations, insomnia, and restlessness. Starting with a low dose and gradually increasing it as appropriate was identified as an effective approach.
Conclusion
This study investigated the attitudes and perceptions of TCM practitioners in Beijing TCM hospitals regarding the dose–efficacy–adverse reaction relationship of E. sinica, providing a reference for the safe and effective clinical use of E. sinica.