2.Immune responses induced by recombinant major outer membrane protein of Chlamydia trachomatis serovar E
Caihong SHENG ; Yuanjun LIU ; Yanfei LI ; Shuping HOU ; Weifeng YAO ; Manli QI ; Quanzhong LIU
Chinese Journal of Dermatology 2009;42(12):817-820
Objective To investigate specific immune responses in mice induced by recombinant major outer membrane protein(rMOMP)of C.trachomatis serovaf E.Methods Thirty-six female BALB/cmice aged 3 to 4 weeks Were divided into three groups.i.e.,adjuvant group vaccinated、with purified rMOMP and Freund's adjutant,solitary group vaccinated with rMOMP only and control group vaccinated with phosphate buffered saline(PBS).All the mice were intramuscularly vaccinated on week 0,2 and 4.Blood samples and vaginal washes were obtained from these mice on week 6,then,mice were challenged with elementary body(EB)of C.trachomatis serovar E at the footpad followed by the observation of delayed hypersensitivity.On week 7.mice were genitally infected with C.trachomatis EB;one week later,blood samples and vaginal washes were obtained again;six weeks later,spleen lymphocytes were isolated from the mice and stimulated bv C.trachomatis or ConA followed by the detection of cell proliferation with MTT assay.In vitro neutralization assay was also performed.ELISA was used to determine the titers of Chlamydia-specific IgO antibody in sera and IgA antibody in vaginal washes,as well as the level of IFN-γ in culture supernatant of lymphocytes and sefa of mice.Vaginal swabs were collected after genital challenge and subjected to C.trachomatis culture.Results The absorbance at 405 ms of Chlamydia-specific IgG antibody and proliferation index of lymphocytes were 0.641±0.059 and 5.085±1.291.respectively,in mice immunized with rMOMP and Frennd's adjuvant.significantly higher than those in mice immunized with rMOMP only(0.424±0.015 and 3.123 ±0.840.both P<0.05).The thickness of right hind footpad increased by 0.324±0.054 mm and 0.272±0.064 mm,respectively,in solitary group and adjuvant group,respectively,with significant difference between the two groups(P<0.05).A significant increase was also observed in the adjuvant group compared with the control group in the above three parameters(all P<0.01).Conclusion The rMOMP of C.trachomatis could efficiently induce Chlamydia-specific humoml and cellular immune responses in mice.
3.Construction of models for persistent genital tract infection with Chlamydia trachomatis in mice with low serum iron
Yong LIU ; Yuanjun LIU ; Weifeng YAO ; Caihong SHENG ; Hui ZHU ; Yan LI ; Quanzhong LIU
Chinese Journal of Dermatology 2010;43(10):702-704
Objective To construct a model for persistent genital tract infection with Chlamydia trachomatis in mice with low serum iron. Methods Iron deficiency was induced in female BALB/c mice by lowiron diet. Mice with or without iron-deficency were inoculated intravaginally with direct instillation of 50 μl of bacterial suspension containing 3.4344 × 107 IFU/ml of C. trachomatis serovar E elementary body 1 week after intramuscular injection of progesterone. Moxifloxacin and ferrous sulfate, alone or in combination were given to inoculated mice with iron deficiency daily for 1 week. Subsequently, vaginal discharge was obtained from mice and observed under an immunofluorescence microscope, and tissue specimens were resected from the uterus,uterine tube and ovary of mice and subjected to pathological examination at various time points. Results Compared with normal-iron mice, iron-deficient mice were infected with C. trachomatis for a longer duration (more than 18 weeks). After 18-week infection with C. trachomatis, there was an obvious chronic inflammation and lymphocytic infiltration in tissues specimens from the uterus,uterine tube and ovary of mice. Immunofluorescent examination of vaginal discharge for C. trachomatis turned negative in 60% of iron-deficient mice treated with both moxifloxacin and ferrous sulfate, but not in any of those receiving moxifloxacin or ferrous sulfate alone. Conclusion As animal models show, low serum iron may contribute to the persistent genital tract infection with C. trachomatis.
4.Cloning, expression, purification and identification of Chlamydia trachomatis polymorphic membrane protein G
Yan LI ; Yuanjun LIU ; Caihong SHENG ; Manli QI ; Weifeng YAO ; Quanzhong LIU
Chinese Journal of Dermatology 2010;43(8):568-571
Objective To clone, express and purify Chlamydia trachomatis polymorphic membrane protein (Pmp G), and to identify its immunogenicity. Methods The Pmp G gene of C. trachomatis serotype E was amplified by PCR, cloned into prokaryotic expression vector PET30a (+). The positive recombinant was transformed into the bacterium E coli (BL-21), identified by enzyme digestion, PCR amplification and gene sequencing. Then, it was induced to express followed by the identification of expression product with SDS-PAGE and Western blotting. The purified protein was used to immunize BALB/C mice to test its immunogenicity. Results PCR produced a 1092 bp-sized DNA fragment, which had a sequence consistent with that of PmpG gene of C. trachomatis E type in the GenBank database. The molecular weight of expression product was 55 kD, which was proved to be the expected size, and Western Blotting confirmed it to be the specific protein. Moreover, special antibodies to PmpG were induced to be generated by mice immunized with the purified protein. Conclusions The constructed prokaryotic expression vector for PmpG is expressed successfully in E. coli, and the expression product shows immunogenicity.
5.Detection of Cd in Chinese Cabbage by Laser Induced Breakdown Spectroscopy Coupled with Multivariable Selection
Hui YANG ; Lin HUANG ; Muhua LIU ; Tianbing CHEN ; Caihong WANG ; Mingyin YAO
Chinese Journal of Analytical Chemistry 2017;45(2):238-244
Heavy metal residue in vegetables is a big concern in the whole world.The aim of this work is to explore the effect of multivariable selection on analyzing Cd in Chinese cabbage polluted in lab by collecting the spectra of laser induced breakdown spectroscopy (LIBS) from the samples.At the same time,the actual Cd content in samples was obtained by anodic stripping voltammetry (ASV).The LIBS spectral range in partial least square (PLS) model was screened by standard normal variable transformation (SNV),first derivative (FD),second derivative (SD) and center treatment (CT) for preprocessing spectra and the optimized method was used for the analysis of interval partial least square (iPLS) and synergy interval partial least square (SiPLS).The results indicated that the method of CT was the best as a comparison with PLS,iPLS and SiPLS.And the intervals of wavelength were 214.72-215.82 nm,215.88-216.97 nm and 225.08 -226.35 nm by utilizing the optimized SiPLS.Here the root mean square error of cross validation (RMSECV) between real content and predicted ones was 1.487,the root mean squared error of prediction (RMSEP) was 1.094,the correlation coefficient (R) was 0.9942,and the average relative error (ARE) was 11.60%.The results displayed that LIBS could predict Cd in vegetables by multivariable selection of SiPLS and the accuracy could meet the requirement of rapid and green analysis of Cd in vegetables.
6.The cross immune responses of the rhesus monkey induced by the E style Chlamydia trachomatis recombinant major outer membrane protein
Yiju LEE ; Weifeng YAO ; Xiaofei ZHAN ; Jing WANG ; Caihong SHENG ; Yanfei LI ; Bin FENG ; Yuanjun LIU ; Manli QI ; Quanzhong LIU
Chinese Journal of Microbiology and Immunology 2012;32(2):161-165
Objective To test cross immune responses induced in rhesus monkeys immunized with the recombinant major outer membrane protein(rMOMP).Methods Six rhesus monkeys were divided into three groups:the group vaccinated with purified rMOMP and Freund's adjutants,the group vaccinated with Freund's adjutants only and the control group vaccinated with PBS.All of the rhesus monkeys vaccinated intramuscularly at 0,2,4 weeks.Two weeks after the last time,The IFN-γand Chlamydia-specific antibody titers in sera,which were determined by ELISA,lymphocyte proliferation assay were performed by MTT,and observ the delayed hypersensitivity and in vitro neutralization assays.Results The result of the monkeys immunized with rMOMP and Freund's adjuvant:the specific immune responses can be observed.The in vitro neutralization and lymphocyte proliferation assays were observed better in the same group.Conclusion After being vaccinated with rMOMP,the monkeys can develop strong and effective Chlamydia-specific cross immune responses.
7.Immune responses induced by the recombinant major outer membrane protein vaccine against Chlamydia trachomatis E serotype in rhesus monkeys
Yiju LEE ; Weifeng YAO ; Caihong SHENG ; Bin FENG ; Xiaofei ZHAN ; Lingjie LI ; Cong YOU ; Yanfei LI ; Ynanjun LIU ; Manli QI ; Quanzhong HU
Chinese Journal of Dermatology 2012;45(7):492-495
[Objective] To observe the specific immune responses induced by the recombinant major outer membrane protein (rMOMP) vaccine against Chlamydia trachomatis E serotype in rhesus monkeys.[Methods] Six rhesus monkeys were equally divided into three groups:adjuvant and protein group vaccinated with purified rMOMP and Freund's adjuvants,adjuvant group immunized with Freund's adjuvants only,and control group immunized with phosphate buffer.All the rhesus monkeys were intramuscularly immunized in the triceps brachii for 3 times at a 2-week interval.Two weeks after the last vaccination,serum,vaginal wash and venous blood samples were collected from the rhesus monkeys,and lymphocytes were isolated from the blood samples.Enzyme linked immunosorbent assay (ELISA) was performed to determine the specific IgG antibody and interferon level in sera and secretory IgA (sIgA) level in wash samples,and methyl thiazolyl tetrazolium (MTT) assay to evaluate the proliferation of lymphocytes after stimulation with Chlaraydia trachomatis serotype E elementary bodies.Delayed hypersensitivity was observed in rhesus monkeys challenged by inactivated Chlamydia trachomatis serotype E elementary bodies.In vitro antibody neutralization assay was conducted with the serum from rhesus monkeys.Indirect immunofluorescenee was used to detect Chlamydia trachomatis in exfoliative vaginal cells from rhesus monkeys from week 1 to 10 after challenge with Chlamydia trachomatis.Data were statistically analyzed by using one-way analysis of variance and least significant difference (LSD) test with the SPSS 14.0 software.[Results] The adjuvant and protein group differed statistically from the adjuvant group and control group in the serum level of specific IgG antibody (1.718 ± 0.213 vs.0.841 ± 0.315 and 0.791 ±0.437,both P< 0.05),interferon ((1086 ± 121.730) ng/L vs.(409 + 53.440) ng/L and (162 ± 48.046) ng/L,both P< 0.05),lymphocyte proliferation index (7.012 ± 1.026 vs.4.473 ± 1.850 and 1A26 ± 1.104,both P<0.01 ) and the diameter of nodus in delayed hypersensitivity assay ( ( 1 1 ± 2.134) mm vs.(3 ± 0.914) mm and 0,both P < 0.01 ).After attack,the exfoliative cells kept positive for Chlamydia trachomatis in the adjuvant and protein group from week 1 to 5,and in the other 2 groups from week 1 to 10,but were negative in the adjuvant and protein group from week 6 to 10.[Conclusion] The rMOMP vaccine can induce a specific,protective,humoral and cellular immune response against Chlamydia tracbomatis in rhesus monkeys.
8.Pathological demography of native patients in a nephrology center in China.
Huiping CHEN ; Zheng TANG ; Caihong ZENG ; Weixin HU ; Qingwen WANG ; Yushen YU ; Xiaodan YAO ; Jianping WANG ; Maoyan ZHU ; Hong ZHOU ; Hong LIU ; Zhihong LIU ; Leishi LI
Chinese Medical Journal 2003;116(9):1377-1381
OBJECTIVETo analysis the pathological demography in Chinese patients undergoing renal biopsy from our nephrology center.
METHODSBetween January 1979 and October 2000 in Jinling Hospital, Nanjing, China, 10,002 attempts of percutaneous renal were performed in patients with renal disease from 33 provinces of China. The pathological classifications were made according to the WHO criteria of 1982 for renal pathology or the modified WHO criteria of 1995 by a panel of pathologists and nephrologists during routine clinical-pathological rounds. The pathological demography between those specimens collected from 1979 - 1989 and those from 1990 - 1999 was compared.
RESULTSThe mean age of the 10,002 subjects undergoing renal biopsy was 31.4 +/- 13.0 years (ranging from 1 to 78 years), with a male to female ratio of 1.3:1; for the 592 renal transplant recipients, the mean age was 37.5 +/- 9.1 years (ranging from 16 to 66 years), with a male to female ratio of 2.36:1. Primary glomerular diseases (PGD) accounted for 71% of the total patients undergoing renal biopsies, secondary glomerular nephritis (SGN) 23%, tubular-interstitial diseases 3.2%, unclassified renal diseases 1.3%, hereditary and congenital renal diseases 1.0%, end stage renal diseases 0.96%, and recently realized or rare renal diseases 0.15%. IgA nephropathy (IgAN) was the most frequent pathological pattern (40%) of PGD, followed by mesangial proliferative lesion (MsPL) (30%), membranous nephropathy (MN) (10%), and focal segmental glomerulosclerosis (FSGS) (6%). Lupus nephritis (LN) was the most pathology common seen (74%) in SGN. During the 22 years of the study period, there was a steady increase in patients with SGN discovered during pathological evaluation of renal disorders. A rise in prevalence was found in IgA nephropathy, MN (both P < 0.001), crescentic glomerulonephritis (P < 0.0001), anti-GBM disease, and hemolytic-uremic syndrome/thrombotic thrombocytopenic purpura related renal damages (both P < 0.001). There was a decrease in endocapillary proliferative glomerulonephritis (P < 0.001) and IgM nephropathy (IgMN) (P < 0.01) from 1990 - 1999 as compared to 1979 - 1989. Infrequent renal pathological entities were also diagnosed in this group, including Niemann Pick disease, Fabry's disease, POEMS syndrome, and lipoprotein glomerulonephropathy.
CONCLUSIONSThis is the largest series of renal biopsy data in China, and therefore may reflect the demographic picture of renal diseases in this country. Changes in prevalence of renal pathological entities were reflected in this group of patients over the last 22 years. In primary glomerular diseases, IgA nephropathy is still the most frequently observed pathological pattern. In SGN, LN appeared the most often. Increased prevalence was found in anti-GBM nephritis and HUS/TTP.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Japan ; epidemiology ; Kidney Diseases ; epidemiology ; pathology ; Male ; Middle Aged ; Prevalence
9.Clinical spectrum of diffuse crescentic glomerulonephritis in Chinese patients.
Zheng TANG ; Yan WU ; Qingwen WANG ; Caihong ZENG ; Xiaodan YAO ; Weixin HU ; Huiping CHEN ; Zhihong LIU ; Leishi LI
Chinese Medical Journal 2003;116(11):1737-1740
OBJECTIVETo investigate retrospectively the incidence, distribution of primary disease and clinicopathologic characteristics of diffuse crescentic glomerulonephritis (DCGN) in Chinese patients.
METHODSOne hundred and seventy-two consecutive patients diagnosed as having DCGN out of 9828 cases of non-transplanting renal biopsies over sixteen years, were studied. DCGN is categorized into three types according to immunopathologic characteristics. The incidence of this disease, its primary diseases, clinical characteristics and serum antineutrophil cytoplasmic antibodies (ANCAs) were analyzed.
RESULTSThe distribution of patients among the three classifications was 8.7% type I, 68.6% type II and 22.7% type III. Clinically, the majority of patients (69.8%) presented rapidly progressive glomerulonephritis (RPGN), but 30.2% manifested a chronic nephritic syndrome or chronic renal failure. In terms of related conditions, 93% were anemic, 61.6% had hypertension, 50.6% oliguria, 45.3% nephrotic syndrome, 43% uremic syndrome and 39.5% displayed gross hematuria. Those patients who were positive in serum for ANCAs had predominantly type III DCGN. Two cases with anti-GBM-antibody crescentic glomerulonephritis and three with lupus nephritis were also positive for ANCAs in serum.
CONCLUSIONDCGN is not rare in Chinese patients. A majority of patients in our study presented with RPGN, but 30.2% manifested a chronic renal failure. Lupus patients with DCGN that were positive for ANCAs had more severe vasculitic lesions.
Adolescent ; Adult ; Aged ; Child ; China ; epidemiology ; Female ; Glomerulonephritis ; classification ; epidemiology ; Humans ; Male ; Middle Aged
10.The effect of hyperbaric oxygen therapy combined with repetitive transcranial magnetic stimulation on patients with cognitive dysfunction after cerebral infarction
Yun REN ; Xudong GU ; Yunhai YAO ; Jianming FU ; Hankui YIN ; Liang LI ; Caihong WU ; Zhongli WANG ; Fang SHEN
Chinese Journal of Physical Medicine and Rehabilitation 2018;40(5):336-339
Objective To observe the effect of hyperbaric oxygen (HBO) therapy combined with repeated transcranial magnetic stimulation (rTMS) on patients with cognitive dysfunction after cerebral infarction.Methods A total of 81 patients with cognitive dysfunction after cerebral infarction were randomly divided into a control group,an HBO group and a combined treatment group,each of 27.In addition to basic medication and traditional rehabilitation therapy,the HBO group was also treated with hyperbaric oxygen,while the combined treatment group received both HBO and rTMS.The mini-mental state examination (MMSE) and the Montreal cognitive assessment (MoCA) were administered before and after the four weeks of treatment.Results The average MMSE scores of the HBO and combined treatment groups were both significantly higher than that of the control group,with the former significantly higher than the latter.Moreover,the average MoCA score of the HBO group was also significantly higher than that of the combined treatment group,and both were significantly higher than that of the control group.Conclusion Hyperbaric oxygen therapy can more effectively improve the cognitive function of patients with cognitive dysfunction after the cerebral infarction when it is combined with repeated transcranial magnetic stimulation.