1.The protective effects of pre-treatment with propoful and Xingnaojing on different characteristic injuries in rat cerebrocortical slices
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2014;26(2):126-130
Objective To investigate the protective effects and mechanisms of propofol and Xingnaojing injection on different characteristic injuries in cerebrocortical slices of newborn rats in vitro. Methods The cerebral cortex of the 7 days-old Sprague-Dawley(SD)neonatal rat was cut into slices. 2,3,5-Triphenyl Tetrazolium(TTC) was used to stain the slices to judge their activities,afterwards they were randomly divided into seven different groups (each,n=12):normal control group(group A),glutamate(Glu)injury group(group B),hydrogen dioxide(H2O2) injury group(group C),propofol preconditioning before Glu injury group(group D),Xingnaojing preconditioning before Glu injury group(group E),propofol preconditioning before H2O2 injury group(group F), Xingnaojing preconditioning before H2O2 injury group(group G). On the 3rd day,the pre-medical treatments or pre-conditionings for D,E,F and G groups were carried out for 24 hours(the concentration of propofol:20 mg/L,the concentration of Xingnaojing:10 μg/mL);the slices were successfully incubated for 4 days,afterwards they were immersed in 1 mmol/L Glu and 0.1 mmol/L H2O2 for 30 minutes respectively to establish the injury models which had no pre-treatment,finally all the groups were transferred into normal cultural medium to incubate till the 7th day. In the above processes,the group A had no specific medical treatment. After all the operations,the changes in brain tissue and cell morphology,the quantity of Nissl body(NISSL)stained by its stain,and the proportion of cells stained with red and green dye after the slices in various groups stained with propidium iodide-acridine orange(PI-AO)were observed,and the apoptosis rate was tested by flow cytometry. Results ①Morphological observation of brain slices:on the 3rd day,the slices appeared mild edematous,3-5 days later,the edema gradually disappeared. Until the 6th day, a large number of typical nerve cells and a few glial cells were seen;on the 7th day,the growth of cells reached the peak,and afterwards,gradually apoptosis played the leading role.②Morphological observation of brain slices stained by hematoxylin-eosin(HE)stain:the neurons of group A presented multilateral-or shuttle-shaped. The cell numbers of groups B and C were significantly lower than the number of group A. In the neuron number sequence,the positions of the numbers of groups D,E,F,G were located in the interval between the number of group A and those of groups B and C,and there were differences in number among groups.③NISSL:Nissl-body of group A could be clearly seen. The numbers of Nissl-body in groups B and C were significantly reduced compared with the number in group A(cell/HP:8.8±2.5,10.3±2.5 vs. 28.9±5.1,both P<0.05). The numbers of Nissl-body in groups D and E were obviously increased compared with the number in group B,and the greater increase being in group D(21.5±4.7 vs. 13.4±3.1, P<0.05). The numbers of Nissl-body in groups F and G were markedly increased compared with the number in group C ,and the greater increase being in group F(23.9±1.9 vs. 19.2±4.1,P<0.05). ④ Double staining with PI-AO:in group A,nearly the total number of nuclei presented fluorescent green in color. In groups B and C,a large number of neurons appeared two types of fluorescence and their edges blurred. The proportions of red staining of neuron cytoplasm in groups D and E were lower than the proportion in group B,and the greater decrease being in group D. The proportions of red staining of neuron cytoplasm in groups F and G were lower than the proportion in group C ,and the greater decrease being in group F.⑤Apoptosis rate:the apoptosis rates of groups B and C were higher than the rate of group A〔(22.00±0.64)%,(21.28±1.44)%vs.(8.57±0.67)%,P<0.05〕;the apoptosis rates of groups D and E were lower than group B,the decrease in group D being greater〔(11.94±0.57)%vs.(18.17±0.65)%,P<0.05〕;the apoptosis rates of groups F and G were much lower than the rate of group C,the decrease in group F being greater〔(10.54±1.24)% vs.(13.12±0.13)%,P<0.05〕. Conclusion The pre-treatment of propoful or Xingnaojing injection has protective effect on Glu or H2O2 injury of in vitro rat cerebrocortical slices,and upon the same injury,the brain protective effect of propoful is more powerful than that of Xingnaojing injection.
2.Effect of 5-HT1 A receptor in hippocampal CA1 region on spatial memory of PTSD rats
Ling LIN ; Guoliang LIU ; Manli SUN
Chinese Journal of Pathophysiology 2017;33(1):98-103
AIM: To investigate the change of long-term potentiation ( LTP ) , and the expression of 5-hydroxytryptamine 1A receptor (5-HT1A receptor) and postsynaptic density protein 95 (PSD-95) in the hippocampus of the rats with posttraumatic stress disorder (PTSD), and to explore the mechanism of 5-HT1A receptor in the regulation of spatial memory in the PTSD rats.METHODS:Healthy adult SD rats (n=36) were randomly divided into control group and mod-el group, with 18 rats in each group.The rats in model group were treated with single prolonged stress to construct the mod-el of PTSD.Morris water maze ( MWM) was used to test the learning and memory ability .The LTP induced by high-fre-quency stimulation (HFS) was detected by electrophysiological method .The protein expression of 5-HT1A receptor and PSD-95 in the hippocampus was determined by Western blot and immunofluorescence .RESULTS: The MWM analysis showed that the latency of the rats searching for the underwater platform in model group was significantly longer than that in control group (P<0.01).The results of electrophysiological analysis showed that the amplitude of the evoked potential in both groups were significantly increased after HFS in the hippocampus , but that in model group was significantly lower than that in control group (P<0.01).The results of Western blot and immunofluorescence analysis showed that compared with control group, the protein expression of 5-HT1A receptor was obviously increased (P<0.05), while the expression of PSD-95 was obviously decreased in model group (P<0.05).CONCLUSION:The spatial memory impairment in the PTSD rats may be associated with the increase in the expression of 5-HT1A receptor and the decrease in the expression of PSD-95 in the CA1 region of hippocampus .
3.Advances in application of flexible bronchoscopy in neonatal intensive care unit
Shaoru HE ; Yumei LIU ; Manli ZHENG
Chinese Pediatric Emergency Medicine 2013;20(1):32-34
Flexible bronchoscopy has become an important diagnostic and therapeutic technique in the neonatal intensive care unit.With the improvement of the instrument and operating techniques of the bronchoscopists,flexible bronchoscopy has been applied in the preterm infant that weighted 600 grams.In this article,a review of application of flexible bronchoscopy in neonates,including diagnostic and therapeutic indications,security in the neonatal applications would be presented.
4.Effect of Curcumine on Expression of Matrix Metalloproteinase- 9 in Human Umbilical Vein Endothelial Cell
Yun ZHANG ; Zongren WANG ; Manli LIU
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(02):-
Objective To investigate the effect of curcumine on the expression of matrix metalloproteinase-9 (MMP-9) in human umbilical vein endothelial cell (HUVEC). Methods HUVEC were cultured and identified in vitro .After cultured with curcumine at different concentration for 24 hours, the expression of MMP-9 in HUVEC was detected by cytoimmunochemical staining method and ELISA . Results All concentrations of curcumine can restrain the expression of HUVEC after co- culturing with the cells for 24 hours. The inhibitory effect got stronger with the increase of the concentration of curcumine. Conclusion The curcumine can depress the expression of matrix metalloproteinase-9 in HUVEC obviously.
5.Detection of serum antibodies to polymorphic membrane proteins (Pmp) in patients with urogenital Chlamydia trachomatis infection
Yan LI ; Yuanjun LIU ; Manli QI ; Caihong SHENG ; Quanzhong LIU
Chinese Journal of Dermatology 2010;43(7):482-484
Objecfive To detect serum antibodies to Pmp in patients with urogenital Chlamydia trachomatis infection and to assess the relationship between Pmp and urogenital C.traehomatis infection.Methods Twenty healthy adults and 77 patients with urogenital C. trachomatis infection were recruited into this study.A 3-month foilow-up was carried out in 43 patients,who were classified into persistent infection group(n=19)and negative-conversion group(n=24).Western-blot was performed to detect serum antibodies to Pmp in all subjects.Results The positivity rate of anti-Pmp antibodies was 90.20% (71/77) in patients,significantly higher than that in the normal controls[20% (4/20),P<0.05].All the 9 types of anti-Pmp antibodies were detected in patients with a varying positivity rates,which were 61.04% (47/77),88.31% (68/77),63.63% (49/77),28.57% (22,77),63.63% (49/77),75.32% (58/77),62.34% (48/77),77.92% (60/77)and 70.13% (54/77) for antibodies against PmpA,PmpB,PmpC,PmpD,PmpE,PmpF,PmpG,PmpH and PmpI respectivelyThe prevalence was highest for anti-Pmp B antibodies and lowest for anti-Pmp D antibodies.There was no significant difference in the positivity rate of anti-Pmp antibodies between persistent infection group and negativeconversion group.Conclusions Anti-Pmp antibodies could be generated in patients infected with C. trachomatis.The immunogenicity of different Pmps is different,and the immunoprotective activity of Pmps is rather weak.Individual differences exist in serum anti-Pmp antibodies among patients.Nine types of Pmps are expressed in patients with urogenital C. trachomatis infection.
6.Immune enhancing effects of interleukin 2 genetic adjuvant on DNA vaccine against Chlamydia trachomatis serovar E
Manli QI ; Jing WANG ; Yuanjun LIU ; Quanzhong LIU ; Jinying CHEN
Chinese Journal of Dermatology 2012;45(5):322-324
ObjectiveTo study the effect of a recombinant plasmid encoding mouse interleukin 2 (mlL-2) on the immunogenicity of DNA vaccine against Chlamydia trachomatis(Ct) serovar E.Methods BALB/c mice were divided into 4 groups to be intramuscularly inoculated with blank plasmid(negative control group),DNA vaccine against Ct serovar E(DNA vaccine group),DNA vaccine against Ct serovar E and a recombinant plasmid containing mIL-2(combination group),and inactivated Ct serovar E elementary bodies (positive control group),respectively.The immunological effects were evaluated by posterior foot pad thickness,proliferation level of spleen lymphocytes,serum level of IL-4 and interferon (IFN)-γ in mice,and the capability to clear Ct genital tract infection.ResultsThe proliferation index of spleen lymphocytes in the combination group and positive control group was similar(3.64 ± 0.41 vs.3.77 ± 0.34),but was significantly different from that in the blank control group and DNA vaccine group (1.37 ± 0.21 and 2.52 ± 0.30).The serum level of IL-4 was(38.49 ± 12.24) pg/ml in the positive control group,significantly higher than in the negative control group,DNA vaccine group and combination group ((25.37 ± 18.93),(24.75 ± 8.49),(21.74 ± 6.43) pg/ml,respectively).With respect to the serum level of IFN-γ,the combination group and positive control group were similar ((1923.3 ± 518.1) pg/ml vs.(2712.5 ± 887.2) pg/ml),but were significantly different from the negative control group and vaccine group((310.8 ± 160.7) pg/ml and(601.3 ± 357.9) pg/ml).Six days after Ct challenge,the exfoliated cells from genital tract were positive for Ct culture in the negative control group,but negative in the other 3 groups.ConclusionIL-2 genetic adjuvant can enhance the immune response,especially Th1 type response,induced by the DNA vaccine against Ct serovar E.
7.Cellular immune responses induced by DNA vaccine against Chlamydia trachomatis E serotype
Manli QI ; Jing WANG ; Quanzhong LIU ; Jinying CHEN ; Naijun TANG
Chinese Journal of Dermatology 2010;43(5):316-319
Objective To study cellular immune responses induced by DNA vaccine against Chlamydia trachomatis (Ct) serotype E. Methods BALB/c mice were divided into three groups to be intramuscularly immunized by blank plasmid (negative control group), DNA vaccine against Ct serotype E (vaccine group), and inactivated Ct elementary body (positive control group), respectively. Two weeks after the last immunization,delayed-type hypersensitivity (DTH) response was evaluated; MTT assay was performed to detect the proliferation of spleen lymphocytes, ELISA to measure the serum level of interferon-γin mice. Some immunized mice underwent a genital challenge with Ct elementary body followed by isolation of Ct from exfoliated epithelial cells in genital tract and pathological examination of cervical tissue from the challenged mice. Results Compared to negative control group, vaccine group and positive control group experienced a stronger DTH response.The lymphocyte stimulating index and serum level of IFN-γwere highest in the positive control group (3.81 ±0.30, 2891.7 ± 1048.8 μg/L), followed by vaccine group (2.35 ± 0.25, 593.3 ± 342.6 μg/L) and negative control group (1.48 ± 0.15, 309.2 ± 157.9 μg/L), and significant difference was observed between the three groups (P < 0.05 or 0.01 ). After Ct challenge, Ct was isolated from exfoliated epithelial cells and cervical tissue was damaged in the negative control group, while in the other two groups, Ct was undetected and genital tract tissue was intact. Conclusions The DNA vaccine against Ct serotype E could induce Ct-specific cellular immune responses to some extent, and offer a protection against vaginal challenge with Ct.
8.Allergic dermatitis to hair dyes: an analysis of 597 cases in Tianjin
Chenwei SUN ; Quanzhong LIU ; Hongjun XU ; Manli QI ; Yumin CHEN
Chinese Journal of Medical Aesthetics and Cosmetology 2011;17(5):359-361
Objective To investigate the morbility and epidemic characteristics of hair dye dermatitis in individuals who dyed their hair in Tianjin.Methods Questionnaires were distributed to the outpatients in the Gerneral Hospital of Tianjin Medical University,students and teachers in Tianjin Medical University,residents in the community and customers in barber shop from Aug.2007 to Mar.2008.The personal data including the methods and site of coloring hair and something correlated to hair dyes were investigated.Results A total of 597 cases with the history of coloring hair were enrolled in the study,including 485 women and 112 men,with mean age of 41 years (ranged from 16- 74years).Among 597 cases,69 cases had allergic reactions to hair dye,including 51 women and 18 men,with mean age of 44 years (ranged from 19-65 years).The median age of the first coloring hair was 40 years (ranged from 3-50 years).The comparison between the sites of coloring hair had no statistic significance (P >0.05),but there was a significant difference between black dyed hairs and col or dyed hairs (P <0.05).Conclusions People with black hair dye are prone to be allergic.
9.Screening of immunodominant proteins for sensitive detection of Chlamydia trachomatis infection by serological test
Xibo GAO ; Meng XIAO ; Jing WANG ; Quanzhong LIU ; Manli QI
Chinese Journal of Microbiology and Immunology 2014;(12):904-907
Objective To detect the antibodies against recombinant chlamydial plasmid-encoded protein 3(rPgp3),chlamydial protease-like activity factor(rCPAF),Ct143 encoded protein(rCT143), Ct101 encoded protein(rCT101),Ct694 encoded protein(rCT694),Ct813 encoded protein(rCT813), Chlamydia membrane protein A(rIncA),Ct875 encoded protein(rCT875),major outer membrane protein (rMOMP)and heat shock protein 60( rHsp60)in serum samples collected form patients with urogenital Chlamydia trachomatis(Ct)infection and to evaluate the antigenicity of those proteins. Methods The re-combinant plasmids expressing the 10 proteins and a blank plasmid were transformed into E. coli BL21 strains,respectively. The transformed E. coli BL21 strains were induced by isopropyl β-D-1-thiogalactopyr-anoside(IPTG)to express recombinant proteins. The glutathione pre-coated 96-well ELISA plates were coa-ted with lysates. Serum samples were collected from 50 patients with Ct infection and 10 patients without Ct infection. ELISA was performed to detect the antibodies against 10 recombinant proteins. Results The anti-bodies against rPgp3,rCPAF,rCT143,rCT101,rCT694,rCT875,rCT813,rMOMP,rIncA and rHsp60 proteins were respectively detected in 44 cases(88% ),38 cases(76% ),37 cases(74% ),36 cases (72% ),33 cases(66% ),31 cases(62% ),30 cases(60% ),26 cases(52% ),24 cases(48% )and 17 cases(34% )out of 50 serum samples. No antibodies against 10 recombinant proteins were detected in the serum samples collected from patients without Ct infection. Conclusion The rPgp3 protein showed the strongest antigenicity among all of the studied proteins,followed by rCPAF and rCT143 proteins. The rHsp60 protein showed the lowest antigenicity.
10.Effects of minocycline on the cognition and expression of BDNF, Bcl-2 and Bax in hippocampus of Alzheimer’s disease model rats
Manli SUN ; Haifeng DENG ; Minghua LI ; Guoliang LIU ; Quanzhong CHANG
Tianjin Medical Journal 2016;44(9):1088-1091
Objective To investigate the effect of minocycline on the cognition and expressions of brain-derived neurotrophic factor (BDNF), apoptosis related factor Bcl-2 and Bax in hippocampus of rats with Alzheimer’s disease (AD).
Methods The rat model was established by microinjection of Aβ25-35 into lateral ventricle. Thirty healthy male SD rats were randomly divided into three groups:control group, model group and minocycline treatment group. Normal saline 1 mL/(kg·d) was intraperitoneally injected in control group and model group. The minocycline treatment group was intraperitoneally injected with minocycline 50 mg/(kg · d) for 14 days. Morris water maze was used to detect the behaviors of animals. The expressions of BDNF, Bcl-2 and Bax in hippocampus were measured by Western blotting and enzyme linked immunosorbent assay (ELISA). The apoptosis of neurons was detected by TdT-mediated dUTP nick-end labeling (TUNEL). Results Minocycline greatly improved the behaviors of AD rats, up-regulated the expressions of BDNF and Bcl-2, and down-regulated the expression of Bax in hippocampus, and reduced cell apoptosis. Conclusion Minocycline plays a protective role in neural function by promoting the growth of neurons and inhibiting the neuronal apoptosis.