1.Expression and Characterization of 21.7 kDa Membrane Protein of Schistosoma japonicum
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(03):-
Objective To express, purify and characterize the 21.7 kDa membrane protein of Chinese strain S. japonicum (SjC21.7). Methods The gene of SjC21.7 was subcloned into the expression vector pGEX 4T 3 to form recombinant plasmid. The recombinant plasmids were transformed into E. coli BL21 and the GST SjC21.7 fusion protein was expressed by IPTG induction. The recombinant SjC21.7 molecule was prepared by affinity chromatography and digested by thrombin. The Kunming strain mice were immunized with the recombinant SjC21.7 molecule to produce anti SjC21.7 antibody. The purified SjC21.7 was recognized by the immunized mouse serum and the sera of rabbits infected by S. japonicum . Results The SjC21.7 gene was subcloned into expression vector pGEX 4T 3, then transformed into E.coli BL21 to express the GST SjC21.7 fusion protein. The recombinant SjC21.7 molecule obtained from the fusion protein could stimulate the mice to produce a high titer of specific antibody and could be recognized by sera of both the immunized and infected rabbits. The sera of immunized mice could also recognize the 21.7 kDa protein molecule of the adult worm antigen (AWA). Conclusion The recombinant and purified SjC21.7 was prepared and showed similar immunological characteristics to the natural SjC21.7 molecule, providing a basis for further investigation of the immunological protection of the recombinant SjC21.7.
2.ULTRA-PATHOLOGICAL STUDY ON THE SYNCYTIUM OF SCHISTOSOMA MANSONI EXPOSED TO CYCLOSPORIN A IN VITRO
Chinese Journal of Parasitology and Parasitic Diseases 1997;0(05):-
Objective] To study the ultra pathological changes of syncytium of Schistosoma mansoni after cyclosporin A (CsA) treatment. [Methods] MF1 mice were infected with Schistosoma mansoni cercariae. Six weeks later, the adult worms were recovered by portal vein perfusion. After the worms were exposed to CsA of 20 ?g/ml for 24 h, the drug induced damage of the worm surface was observed by SEM and TEM. [Results] Incubation of male and female schistosomes with 20 ?g/ml of CsA for 24 h resulted in disruption of the tegument and rupture of the spines. Progressive surface damage and swelling and vacuolization of the tegument led to eventual disruption of the syncytium. [Conclusion]The antischistosomal action of CsA is direct, the syncytium is the main site for CsA attack.
3.Fusion Expression and Antigenicity Analysis of MiracidialAntigen from Eggs of Schistosoma japonicum
Jianfa LIU ; Chuanxin YU ; Yinchang ZHU ; Xuren YIN ; Yongliang XU
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(03):-
Objectives To express the miracidial antigen from eggs of Schistosoma japonicum (Chinese mainland strain) (SjMP10), and investigate the role of the miracidial antigen during the hepatic granuloma formation of schistosomiasis. Methods A pair of specific primers was designed and synthesized according to the nucleotide sequence of the open reading frame of the miracidial antigen gene. The open reading frame of the miracidial antigen gene was amplified, digested by restrictive enzyme(BamHI, SalI), and cloned directly into the expression plasmid pGEX-4T-3 to construct the recombinant plasmid. The recombinant plasmids were transformed into E. coli BL21(DE3), and induced by IPTG to express the fusion protein of GST-SjMP10. The expressed fusion protein was purified by electric elution method, and its antigenicity was examined by Western blotting and lymphocyte proliferation test. Results The gene of miracidial antigen was cloned into the expression plasmid pGEX-4T-3. After induced by IPTG, the recombinant expressed a fusion protein of GST-SjMP10, with a molecular weight of 39 000 approximately. The purified fusion protein showed proper antigenicity that could be recognized by the sera of rabbits heavily infected by Schistosoma japonicum and could stimulate the proliferation of splenic lymphocytes of infected BALB/c mice. Conclusion The miracidial antigen from eggs of Schistosoma japonicum was expressed successfully.
4.Expression of jumonji domain-containing histone demethylase 2 and estrogen-related receptor alpha in postmenopausal osteoporosis
Hongyu TANG ; Lujue DONG ; Shaochuan HUO ; Cheng GUO ; Chi ZHOU ; Jianfa CHEN ; Yong LIU ; Haibin WANG
Chinese Journal of Tissue Engineering Research 2016;(2):167-172
BACKGROUND:Jumonji domain-containing histone demethylase (JMJD) can promote osteoblast differentiation, and estrogen-related receptor alpha (ERRα) can promote osteoblast differentiation and increase bone formation. However, little is reported on the association between postmenopausal osteoporosis andJMJD and ERRα. OBJECTIVE: To study the changes in the JMJD2 family expression in patients with postmenopausal osteoporosis. METHODS: Postmenopausal patients with osteoarthritis of the hip scheduled for total hip arthroplasty, aged 50-70 years, were enroled, including 10 postmenopausal osteoporosis patients (experimental group) and 10 patients with no postmenopausal osteoporosis (control group). During the arthroplasty, the cancelous bone specimens from the femoral head were colected. Then, immunohistochemistry and western blot assay were used to detect expression of histone demethylase (JMJD2A, JMJD2B), histone methylation (H3K9me3, H3K36me3) and ERRα. RESULTS AND CONCLUSION:In the experimental group, the expressions of JMJD2A, JMJD2B and ERRαwere from weakly positive to positive; these expressions were significantly lower in the experimental group than the control group (P < 0.05). The expressions of H3K9me3 and H3K36me3 were significantly higher in the experimental group than the control group (P < 0.05). These findings indicate that the expression of JMJD2A and JMJD2B is consistent with the expression of ERRα in the patients with postmenopausal osteoporosis, and JMJD is likely to serve as an antagonistic enzyme of osteoporosis.
5.Epidemiological Investigation on the Intermediate Hosts of Paragonimus in Ninghai County of Zhejiang Province
Jianfa LIU ; Yan WU ; Zhiyuan TANG ; Dan MENG ; Shujuan YANG ; Wenwen JIANG
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(04):-
Freshwater crabs and snails were collected from Ninghai County in Zhejiang Province, and examined respectively for Paragonimus metacercariae and cercariae. Among 97 freshwater crabs found, the prevalence was 11.3% (11/97) with a mean intensity of 1 metacercariae per crab. It was 10.2% (5/49) and 20.2% (4/20) in the groups weighted 5-15 g and 15-25 g respectively, with an average intensity of 1, and no metacercariae were found in weight group of 25-35 g. Two positive crabs were found from 20 crabs with a low weight (
6.Study of correlation between biochemical markers of bone metabolism and postmenopausal osteoporotic vertebral fractures
Shaochuan HUO ; Lujue DONG ; Hongyu TANG ; Yong LIU ; Hai GUO ; Jianfa CHEN ; Zhangrong DENG ; Delong CHEN ; Haibin WANG
Chongqing Medicine 2017;46(1):48-50
Objective To study correlation between biochemical markers of bone metabolism and postmenopausal osteoporot-ic vertebral fractures.Methods The clinical data of 100 cases with postmenopausal osteoporotic were study retrospectively.Fifty patients were postmenopausal osteoporotic,the rests were postmenopausal osteoporotic vertebral fractures.Lumbar spine,hip BMD,serum P1NP,β-CTX,N-MID,25-(OH)VitD and Ca2 + were recorded.Results There was a significant difference among ser-um P1NP,β-CTX and 25-(OH)VitD(P <0.05 ).There was positive correlation between postmenopausal osteoporotic vertebral fracture with serum P1NP (P <0.05),and negative correlation with serum 25-(OH)VitD (P <0.05),but had no correlation with serumβ-CTX (P >0.05).Conclusion Serum P1NP and 25-(OH)VitD could predict risk of postmenopausal osteoporotic vertebral fractures.Biochemical markers of bone metabolism combined with BMD could reduce postmenopausal osteoporosis fractures.
7.Clinical analysis on the surgical treatment of sacrococcygeal tumors
Bin HU ; Bo HU ; Jianfa NI ; Deheng LI ; Guangxin ZHOU ; Xiaozhou LIU ; Chengjun LI ; Meng LU ; Xin SHI ; Sujia WU
Journal of Medical Postgraduates 2014;(4):387-389
Objective The incidence rate of sacrococcygeal tumor is low , however , due to its special position , clinical symptoms are inclined to be more severe .The aim of the article was to explore the methods and therapeutic effects of operative treat-ment for sacrococcygeal tumors . Methods A retrospective analysis was made on 44 cases of sacrococcygeal tumors treated in our hospital from January 2008 to December2012 .Of all the cases , there were 11 chordomas , 9 neurinoma , 4 malignant fibrous histioto-ma, 4 giant cell tumor, 7 metastases tumors and 9 others.All the patients had definite pathological diagnosis after operation .Operative treatments involved simple resection of rumors , simple resection of lesions and resection of lesions plus screw-rod system internal fixa-tion. Res ults No patients died in perioperative period .Follow-up was made in all patients for the average time of 3.3 years(4 months to 59 months).Except for 4 patients'delayed healing of incision operation and 1 patient′s healing of operation incision after de-bridement , all the other patients healed after enhancing the wound dressing , among which there are 3 cases of chordoma recurrence , 1 case of giant cell tumor recurrence , 3 death cases of malignant fibrous histiotoma in 2 years and 1 case alive with tumor .All patients with metastases tumor died in 3 years.Except 1 patients with osteosarcoma alive with tumor , all patients′symptoms had been alleviated after operation. Conclusion Chordoma and giant cell tumor still have a high recurrence rate after operation . The extent of rumor resection and nerve preservation are determined by the range of tumors.Preoperative embolization of the internal iliac artery can clearly reduce bleeding and improve the operation safety .
8.In vitro differentiation of human skin-derived mesenchymal stem cells into lymphocytes: Possibility evaluation
Liping GUAN ; Jie YU ; Bing HUANG ; Ting LUO ; Jianfa HUANG ; Qian LIU ; Liping LIN ; Min ZHANG ; Kaijing LI ; Xigu CHEN
Chinese Journal of Tissue Engineering Research 2010;14(19):3601-3605
BACKGROUND: Previous research has demonstrated that dermal tissue has mesenchymal stem cells, which have a possibility of autologous transplantation. If the mesenchymal stem cells derived from the skin differentiate into lymphocytes under a certain condition, the immune system disease can be solved generally.OBJECTIVE: To investigate the possibility of differentiation of human skin-derived mesenchymal stem cells into lymphocytes. METHODS: Surface marker expression was detected in the 14th passage human skin-derived mesenchymal stem cells using flow cytometry. Transdifferentiation medium of human skin-derived mesenchymal stem cells consisted of human lymphocyte supernatant and fresh human skin-derived mesenchymal stem cells based on the ratio of 7:3. Inverted microscope was employed to observe morphological changes, and flow cytometry was used to detect surface marker expression in the lymphocytes at 1-8 days after induction. Self-marker expression of human skin-derived mesenchymal stem cells was then detected at 3,6, and 9 days after induction.RESULTS AND CONCLUSION: Human skin-derived mesenchymal stem cells stably expressed self-specific marker CD73, Vimentin and so on, but did not express specific markers of hematopoietic system, I.e., CD34, CD45 and so on, lowly expressed HLA-I, but did not express HLA-DR at all. At 3 days after induction, the cell volume significantly increased, cell proliferation rate was significantly lower than before induction, and a lot of cystic-like particles with strong refraction were observed in or between cells. The CD45 lymphocyte expression was not significantly changed, but CD3, CD19, CD16, CD4, and CD8 expression rates of human skin-derived mesenchymal stem cells were linearly increased at 1-4 days after induction and stabilized at 5-8 days after induction. In addition, CD37, CD34, Vimentin, and HLA-DR expressions were not changed at 3, 6, and 9 days after induction, but HLA-I expression rate was gradually increased with the prolongation time of induction. This suggested that human skin-derived mesenchymal stem cells can differentiate into lymphocyte and potentially participate in repairing immune system injury.
9.Risk factors for heterotopic ossification following total hip arthroplasty in patients with ankylosing spondylitis
Yong LIU ; Shaochuan HUO ; Chi ZHOU ; Hongyu TANG ; Delong CHEN ; Jianfa CHEN ; Hai GUO ; Zhangrong DENG ; Haibin WANG
Chinese Journal of Tissue Engineering Research 2017;21(11):1641-1646
BACKGROUND: Heterotopic ossification (HO) is common following primary total hip arthroplasty (THA) in patients with ankylosing spondylitis (AS), which may cause certain influence on functional recovery.OBJECTIVE: To explore the risk factors for HO after primary THA in AS patients.METHODS: The clinical and radiological data from 87 patients (132 hips) with AS undergoing primary THA between June 2011 and December 2015 were retrospectively analyzed, and followed up for more than 6 months. The radiological information included preoperative and postoperative hip anteroposterior and lateral radiographs. The presence of HO surrounding the prosthesis was evaluated on the radiographs at the last follow-up and graded according to the Brooker classification. Risk factors for HO were divided into invariable factors (age, sex, course and with or without ankylosed hip) and variable factors (preoperative C-reactive protein level, preoperative erythrocyte sedimentation rate, intraoperative blood loss, operation time, prosthesis types and anesthesia methods) to determine the pertinent risk factors.RESULTS AND CONCLUSION: (1) Totally 43 hips (32.6%) were found to have developed into HO. (2) Invariable risk factors including male (P=0.029), preoperative ankylosed hip (P < 0.001), and course (P=0.029) increased the prevalence of HO. Among the variable risk factors, prolonged operation time (P=0.031) and general anesthesia (P=0.003)were associated with the increased occurrence of HO. Age, preoperative C-reactive protein level and erythrocyte sedimentation rate, intraoperative blood loss, and prosthesis types had no obvious correlation with HO. (3) These results suggest that to prevent the formation of HO following THA in AS, efforts to reduce the operation time and avoid general anesthesia should be considered.
10.Relationship between thyroid function and cardiovascular diseases
Mei LIU ; Bingfeng ZHOU ; Youwei SHI ; Shaodong XU ; Ying MENG ; Jianfa ZHENG ; Qinghe XU ; Minyu LIU ; Ying GAO ; Fei GAO ; Wei LIU
Chinese Journal of cardiovascular Rehabilitation Medicine 2015;24(3):267-270
Objective:To explore the relationship between thyroid function and cardiovascular diseases .Methods:A total of 980 patients undergoing thyroid function examination during hospitalization were selected . According to their thyroid function ,they were divided into normal thyroid function group (normal group ,n= 930) , hyperthy-roidism group (n=18) ,and hypothyroidism group (n=32) .Clinical data were analyzed ,blood lipids and coagula-tion function indexes were examined and compared among three groups . Results:Compared with normal group ,the incidence rate of atrial fibrillation significantly rose ,incidence rate of hypertension ,cardiac insufficiency signifi-cantly reduced;levels of TC、TG、 LDL-C、 HDL-C significantly reduced ,activated partial thromboplastin time significantly extended in hyperthyroidism group , P<0.01 all;incidence rate of coronary heart disease significantly rose ,levels of TG、HDL-C significantly rose ,levels of TC 、LDL-C significantly reduced in hypothyroidism group , P<0.01 all;Compared with hyperthyroidism group ,the incidence rate of hypertension ,coronary heart disease sig-nificantly rose ,atrial fibrillation significantly reduced ,P<0.05 or <0.01 ;levels of TG、LDL-C、HDL-C signifi-cantly rose ,TC level significantly reduced in hypothyroidism group , P<0. 01 all .Conclusion:Thyroid function is closely related to cardiovascular diseases .so it′s suggested that thyroid function detection should be regarded as a routine examination in patients with cardiovascular diseases and a follow-up index for those with thyroid dysfunc-tion .