1.Gene cloning, optimized expression and immunogenicity evaluation of tetanus toxin fragment C.
Ming-yong WANG ; Ya-ni ZHANG ; Ming LEI ; Da-ming ZUO ; Li-yun ZHANG ; Zheng-liang CHEN
Journal of Southern Medical University 2008;28(5):731-735
OBJECTIVETo obtain highly purified tetanus toxin fragment C (TTC) with good immunogenicity.
METHODSThe gene fragment encoding TTC was amplified from Clostridium tetani plasmid DNA by PCR, inserted into the vector pET43.1a (+) and expressed in E. coli BL21(DE3)plysS. After purification using Ni2+-chelate affinity chromatography, the expressed fusion protein was digested by thrombin and the resultant TTC protein was purified with Ni2+-chelate affinity chromatography followed by identification with SDS-PAGE and Western blotting. The purifed TTC protein was then used to immunize mice to test its immunogenecity.
RESULTSThe 1373-bp gene fragment encoding TTC was obtained, and the constructed recombinant expression vector pET43.1a (+)-TTC was successfully expressed in E. coli BL21(DE3)plysS. SDS-PAGE identified a recombinant fusion protein with relative molecular mass (Mr) of 117 000, which accounted for 22% of the total bacterial protein. The TTC protein with Mr of 50 000 was obtained after purification of the thrombin digestion products of the fusion protein, with a purity reaching 95.5%. Both the fusion protein and TTC protein could be recognized by anti-tetanus toxin antibody as shown by Western blotting. The titer of the anti-serum from mice immunized with the TTC protein was 1:25 600, and the anti-serum could specifically bind to tetanus toxin.
CONCLUSIONHighly purified and immunogenetic TTC protein has been successfully obtained, which provides a good model antigen for studying antigen presentation and immune responses in vivo.
Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Peptide Fragments ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Tetanus Toxin ; biosynthesis ; genetics ; immunology ; Tetanus Toxoid ; immunology
2.Incidence of seroresistance of syphilis and its relevant factors.
Lin-Na WANG ; Ya-Gang ZUO ; Yong-Xin LIU ; Xiu-Rong LIU ; He-Yi ZHENG
Acta Academiae Medicinae Sinicae 2008;30(3):338-341
OBJECTIVETo investigate the incidence of seroresistance of syphilis and analyze its relevant factors.
METHODSThe clinical data of 131 patients with syphilis were retrospectively analyzed. The incidence of seroresistance was investigated and the correlation between seroresistance and factors including age, gender, original titer, disease course, and medications were analyzed.
RESULTSThe incidence of seroresistance was not significantly different among patients with different ages and genders, but was significantly different among patients with different disease courses, antibody titers, and medications. Patients with a baseline serum rapid plasma reagin (RPR) titer of greater than 1: 8, a latent course, or a macrolide therapy history had higher incidences of seroresistance (i. e., 61%, 45.6%, and 72.7% respectively) than those who had a lower RPR titer, a primary course, or a benzathine penicillin therapy history.
CONCLUSIONSyphilis patients with a high baseline RPR titer, a latent course, and a macrolides therapy are prone to be seroresistant.
Adolescent ; Adult ; Aged ; Female ; Humans ; Macrolides ; therapeutic use ; Male ; Middle Aged ; Reagins ; blood ; Syphilis ; blood ; diagnosis ; drug therapy ; immunology ; Syphilis Serodiagnosis ; methods ; Young Adult
3.Analysis of 1125 syphilis cases.
Jun LI ; Lin-na WANG ; He-yi ZHENG ; Ya-gang ZUO ; Yong-xin LIU ; Xiu-rong LIU
Acta Academiae Medicinae Sinicae 2010;32(2):185-189
OBJECTIVETo summarize the epidemiological, clinical and serological features of syphilis.
METHODThe clinical data of 1125 patients with syphilis were retrospectively analyzed.
RESULTSNon-marital sexual intercourse is the main route of infection. The 129 cases of primary syphilis were featured by extragenital chancres and the 357 cases of secondary syphilis were featured by scaling macules on the palms and soles and condylomata lata. Early latent syphilis were detected in 178 cases, late latent syphilis in 174 cases, and latent syphilis of unknown duration in 285 cases. Concurrent infection with other sexually transmitted diseases were noted in 107 cases (9.51%). Most patients were treated with benzathine penicillin, and only 31 patients (2.75%) were allergic to this therapy. Except for the primary syphilis, the sensitivity of syphilis serological tests (including rapid plasma regain test, treponema pallidum particle agglutination, treponema pallidum hemagglutination assay, and fluorescent treponemal antibody-absorption) were higher than 97%.
CONCLUSIONSThe incidence of latent syphilis is high, while the incidence of late syphilis is low. Penicillin remains the treatment of choice for syphilis.
Adult ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Syphilis ; diagnosis ; epidemiology ; therapy ; Young Adult
4.Mutation Analysis of CIB2 196C>T 272T>C and 297C>G in Patients with Non-syndromic Hearing Impairment
Qi LI ; Jun-Dong HE ; Jin-Li WANG ; Yong-Qin ZHENG ; Rong-Xia ZUO ; Ya-Lian SA
Journal of Kunming Medical University 2018;39(5):7-10
Objective To investigate the gene mutations of calcium-and integrin-binding protein 2 (CIB2) 196C>T, 272T > C and 297C > G carried by students with non-syndromic hearing impairment from special educational schools in Yunnan Province. Methods The experimental group included 337 students with non-syndromic hearing impairment who failed to carry deafness gene with GJB2 (35 del G, 176_191 del 16,235delC, 299_300 del AT), GJB3 (C538T,G547A), mtDNA 12S rRNA (A1555G, C1494T), and SLC26A4 (IVS7_2A>G, A2168G) . The control group consisted with 150 healthy people. Genomic DNA was isolated from peripheral blood with EDTA anti-coagulate. The subject's DNA fragments including CIB2 196C>T, 272T > C and 297C> G were amplified by polymerase chain reaction (PCR), and subsequently analyzed by direct sequencing to identify deafness-associated mutations. Results Both in the experimental group and control group, we failed to find the mutation of CIB2 196C>T, 272T>C and 297C>G in all individuals. Conclusion Mutations in CIB2 gene 196C>T, 272T>C and 297C>G are not a frequent cause of non-syndromic hearing loss among deaf people in Yunnan province. It provided important information for deafness with formulating landscape of gene screening in this region.
5.Purification of plasmin-alpha2-antiplasmin complex by lysine-sepharose 4B affinity chromatography employing sequential elutions.
Jian-Xin LI ; Yun ZHOU ; Hong WANG ; Zuo-Ya ZHENG ; Dao LI ; Hong-Li WANG
Journal of Experimental Hematology 2004;12(3):355-358
To easily obtain plasmin-alpha(2)-antiplasmin complex (PAP) with high purity, the purification procedure was improved by authors. After urokinase activated fresh human platelet-deficient plasma had been applied to Lysine-Sepharose 4B affinity chromatography column, elutions were sequentially performed by several eluents with different ingredients. The results showed that 3.0 mg PAP could harvested from 100 ml fresh plasma by this method and the whole procedure could be finished within several hours. In conclusion, this procedure is simple, rapid, economical and high-yield.
Blotting, Western
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Chromatography, Affinity
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Electrophoresis, Polyacrylamide Gel
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Fibrinolysin
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analysis
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isolation & purification
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Humans
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Sepharose
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analogs & derivatives
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alpha-2-Antiplasmin
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analysis
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isolation & purification
6.Pharmacokinetics of omeprazole in Chinese Muslim and Han nationalities healthy volunteers after a single dose administration
Tao GUO ; Zuo-Ming SUN ; Dong-Ya XIA ; Yi ZHENG ; Yong XIAO ; Ning-Qin WANG ; Hong-Liang GUO ; Peng LIU
The Chinese Journal of Clinical Pharmacology 2010;26(2):110-113
Objective To study the pharmacokinetics of omeprazole tablets in Chinese Hui and Han healthy volunteers.Methods Twenty volunteers were involved in the study,including ten Chinese Hui volunteers and ten Han volunteers.Each healthy volunlteers were given asingle oral dose(40 mg)of omeprazole tablet.The concentration of omeprazole in plasma was determined by RP-HPLC and the pharameters were calculated by DAS 2.0 software.Results The main pharmacokinetic parameters of Chinese Hui and Han were as follows:C_(max) were (941.94±446.08) and (760.49±581.23) μg·L~(-1);t_(max) were(2.7±0.68)and(2.70±0.82)h;t_(1/2) were(2.43±2.83)and(1.60±1.28)h;AUC_(0-12) were (2.29±1.12)and(1.44±7.98)mg·h·L~(-1);AUC_(0-∞) were (2.33±1.11)and(1.47±7.69)mg·h·L~(-1),respectively.Conclusion Individual difference is significant in C_(max),AUC_(0-12),AUC_(0-∞) after an oral omeprazole,but the pharmacokinetic parameters obtained from the results of ststistical analysis show no significant difference in the main pharmacokinetic of omeprazole between Chinese Hui and Han.
7.A new classification of nevus of Ota.
Wen-hui HUANG ; Hong-wei WANG ; Qiu-ning SUN ; Hong-zhong JIN ; Yue-hua LIU ; Dong-lai MA ; Ya-gang ZUO ; He-yi ZHENG ; Kuo WAN ; Quan JING ; Yong-liang ZHAO
Chinese Medical Journal 2013;126(20):3910-3914
BACKGROUNDThe nevus of Ota, is a common benign pigmentary dermatosis, mainly involve innervation area of first and second branch of trigeminal nerve. The classification of nevus of Ota was proposed by Tanino, based on 26 cases of nevus of Ota from 1937 to 1940. Studies about its classification are rarely seen in last 70 years, while it is still practical today.
METHODSBased on the clinical photographs, 1079 consecutive patients with nevus of Ota were verified and reclassified according to the innervation areas of the trigeminal nerve branches.
RESULTSIn these 1079 cases, 866 patients were in line with Tanino's classification (80.26%), and 213 patients were not (19.74%). We put forward a new clinical classification (Peking Union Medical College Hospital classification, PUMCH classification) of nevus of Ota based on the innervation area of the trigeminal nerve branches, composed of 5 types and 14 subtypes. The 5 types were as follows: Type I - pigmentation maculeses involving the innervation area of one of the three trigeminal nerve branches, of which there were 424 cases (39.3%), comprising 6 subtypes; Type II - pigmentation macules involving the innervation area of two branches of the three trigeminal nerve branches, of which there were 221 cases (20.48%), comprising 4 subtypes; Type III - pigmentation macules involving the innervation area of all three trigeminal nerve branches, of which there were 361 cases (33.45%), comprising 2 subtypes; Type IV - bilateral type, in which the pigmentation macules involves the bilateral cheek, of which there were 63 cases (5.84%), comprising 2 subtypes; and Type V - complications occurred in the patient, of which there were 10 cases (0.93%).
CONCLUSIONThe new classification of nevus of Ota is based on the innervation area of the trigeminal nerve branches, and it covers all types of Tanino's classifications; on that basis, some new types and subtypes are brought in and cover almost every clinical condition.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Middle Aged ; Nevus of Ota ; classification ; diagnosis ; Trigeminal Nerve ; pathology ; Young Adult
8.Retrospective analysis of 50 cases of senile syphilis.
Lin-na WANG ; He-yi ZHENG ; Ya-gang ZUO ; Jun LI
Acta Academiae Medicinae Sinicae 2014;36(2):198-200
OBJECTIVETo summarize the epidemiological features of senile syphilis.
METHODThe clinical data of 50 elderly (>60 years) patients with syphilis who were treated in the Clinic of Sexually Transmitted Diseases of Peking Union Medical College Hospital were retrospectively analyzed.
RESULTSThere were 31 men and 19 women (male to female ratio:1.6:1). Non-marital sexual intercourse was the main route of transmission. The disease was mainly found before surgeries. Among these 50 syphilis cases, 3 were primary syphilis, 13 were secondary syphilis, 10 were early latent syphilis, 12 were late latent syphilis, and 12 were latent syphilis of unknown duration. Co-morbidities with other sexually transmitted diseases were found in 4 patients. The rate of sero-resistance was 40%.
CONCLUSIONSThe incidence of syphilis in the senile population is higher than we expected. Routine serological screening for syphilis is required in clinical settings. The underlying diseases, human hosts-related spirochetes, and laboratory testing techniques are important reasons for the false positive findings in senile patients.
Aged ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Syphilis ; classification ; diagnosis ; epidemiology
9.Analysis of influenza vaccination coverage, recommendation behaviors and related factors among health care workers in Nanshan district of Shenzhen city under the free policy between 2019 and 2020.
Shi Qiang JIANG ; Yu Wei CAI ; Ran ZUO ; Li Fang XU ; Jian Dong ZHENG ; Hao Ya YI ; Zhi Bin PENG ; Luzhao FENG
Chinese Journal of Preventive Medicine 2022;56(11):1565-1570
Objective: To investigate the current situation of influenza vaccination, vaccination willingness, recommended behavior and influencing factors of health care workers (HCWs) under the policy of free vaccination. Methods: A cross-sectional survey was conducted among 3 167 medical staff from 8 hospitals in Nanshan district of Shenzhen city based on a web-based questionnaire platform. The logistic regression was used to analyze the data. Results: The influenza vaccination rate in HCWs was 23.97%, and the recommendation rate was 25.69% from 2019 to 2020. Staff with high professional titles, high academic qualifications, and positive awareness about influenza vaccine had a higher vaccination rate. The main reasons for not recommending influenza vaccine were the fear of patients' misunderstanding of commercial benefits, fear of possible disputes caused by recommended vaccination, lack of national or institutional requirements for recommended influenza vaccine, and fear of adverse reactions of influenza vaccines. Conclusion: Under the free policy, the influenza vaccination rate and recommendation rate of HCWs in Nanshan district of Shenzhen city are relatively low. Strengthening health education on influenza and related knowledge, publicizing the policy of free influenza vaccination, providing convenient vaccination services and promoting the construction of relevant policies and regulations are the key to improve the influenza vaccination rate and recommendation rate among HCWs.
Humans
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Influenza Vaccines
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Influenza, Human/prevention & control*
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Vaccination Coverage
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Cross-Sectional Studies
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Attitude of Health Personnel
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Vaccination
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Health Personnel
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Surveys and Questionnaires
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Policy
10.Efficacy of Nonmyeloablative Allogeneic Hematopoietic Stem Cells for 14 Case of Severe Acquired Aplastic Anemia.
Shao-Fei WANG ; Ya-Jing HUANG ; Chang-Lin YU ; Qi-Yun SUN ; Jian-Hui QIAO ; Kai-Xun HU ; Hong-Li ZUO ; Zheng DONG ; Zhi-Qing LIU ; Yi WANG ; Xiao-Mei HUANG ; Hui-Sheng AI ; Mei GUO
Journal of Experimental Hematology 2015;23(4):1097-1102
OBJECTIVETo investigate the therapeutic efficacy of nonmyeloablative allogeneic hematopoietic stem cells transplantation for severe acquired aplastic anemia (SAA).
METHODSFourteen patients with severe acquired aplastic anemia received nonmyeloablative allogeneic hematopoietic stem cells transplantation from HLA matched sibling donors, among them 8 cases were dagnosed as SAA-I, 6 cases were diagnosed as SAA-II. The conditioning regimen consisted of fludarabine (FIUD), cyclophosphamide (CTX) and anti-thymocyte globulin (ATG/ALG). The prophylaxis for graft-versus-host disease (GVHD) was performed with cyclosporine (CsA) combined with mycophenolate mofetil (MMF) or tacrolimus (FK506).
RESULTSAll the patients gained a quick successfully engraftment of donor hametopoietic cells. The mean recovery time for neutrophil and platelet was 9 d and 13 d respectively. All the patients have acquired a full donor chimerism before 14 d. There were only 2 cases of GVHD: one out of them was acute skin GVHD (grade I) at day 70 after transplantation and the other was chronic liver GVHD (grade I) in 1 years after transplantation, the GVHD more than degree II did not coccur in all patients, 9 patients with bacterial and fungal mixed infection and (or) virus infection were observed, and improved after anti-infection therapy. The median follow-up time were 54.5 months (ranged between 5-144 months), and 12 patients remain disease-free survival currently, only 2 patients died of fungal infectin.
CONCLUSIONTransplantation of nonmyeloablative allogeneic hematopoietic stem cell is safe and effective for the treatment of severe acquired aplastic, but the prevention, treatment and monitoring of infection need to be enhance.
Allografts ; Anemia, Aplastic ; Antilymphocyte Serum ; Cyclophosphamide ; Cyclosporine ; Disease-Free Survival ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; Humans ; Mycophenolic Acid ; analogs & derivatives ; Neutrophils ; Siblings ; Tissue Donors ; Transplantation Conditioning ; Vidarabine ; analogs & derivatives