2.Inhibitory effect of taurine on lens epithelial cell apoptosis
Wenjuan LUO ; Chuanfu WANG ; Hui LI ; Ju KANG ; Honglu YAN
Chinese Journal of Tissue Engineering Research 2008;12(11):2197-2200
BACKGROUND:Taurine is an important non-enzymatic system antioxidant in the lens.The mechanism of its anti-oxidative effect is mainly to protect lens from oxidative injury by anti-lipid peroxidation.OBJECTIVE:This study was to observe the effect of exogenous taurine on lens epithefial cell apoptosis-induced by H2O2 in vitro.DESIGN:A randomized controlled animal experiment.SETTING:Department of Ophthalmology,Affiliated Hospital of Qingdao University Medical College.MATERIALS:Seventy-five adult New Zealand standard tabbits,of either gender,weighing 1.5-2.5 kg,were provided by Qingdao Laboratory Animal Center.Reagent kit for in situ detecting cell apoptosis(Sigma Company,USA),taurine and H2O2(Shanghai Guangda Chemical Reagent Factory,China)were included in this study.METHODS:This study was performed in the Department of Ophthalmology and Central Laboratory.Affiliated Hospital of Qingdao University Medical College from Mav 2005 to June 2007.Rabbit lenses were harvested and randomly divided into 3 groups:control group,in which,clear lenses were incubated in non-serum and non-phenolsulfonphthalein MEM medium which was renewed every 24 hours,H2O2 group,in which,clear lenses were incubated in non-serum and non-phenolsulfonphthalein MEM medium containing 1 mmol/L H2O2 with addition of 62 μL H202(30 g/L)every 6 hours,and H2O2+taurine group,in which,clear lenses were incubated in non-serum and non-phenolsulfonphthalein MEM medium containing 1 mmol/L H2O2 and 10 g/L taurine.which was renewed every 6 hours.The protocol was conducted in accordance with ethical guidelines for the use and care of animals.MAIN OUTCOME MEASURES:Observation of lens opacity 6.12,24.48 and 72 hours after culture;Lens epithelial cell apoptosis determined by DNA in situ end labeling and DNA fragment analysis.RESULTS:Lens opacity:The lens opacity in the H2O2 group was aggregrated gradually along with the time of oxidative injury.The lens opacity in the H2O2 group was severer than that in the H2O2+taurine group.Lens epithelial cell apoptosis:There were no apoptotic cells in the control group within 72 hours.The number of apoptotic cells in the H2O2 group was increased gradually with the prolonged time of oxidative injury.Till the 72nd hour,the cells were all tamed into apoptotic cells.A few aopototic celIs were found in the H2O2+taurine group since hour 24,and then were more and more,and the number of apoptotic cells accounted for about 30%at hour 72.The apoptotic rate in the H202+taurine group was significantly lower than that in the H2O group at each time point(q=8.6845,P<0.01).There was no significant difference in apoptotic rate between the H202+taurine group and the control group (P>0.05). Findings of DNA fragmentation assay:The DNA"ladder"was found in the H2O2 group at hours 24,36,48 and 72,while no DNA"ladder"but only normal electrophoresis straps were present in the other two groups 24 hours after culture.CONCLUSION:Taurine can inhibit the oxidative injury-induced apoptosis of rabbit lens epithelial cells,and alleviate lens opacity.
3.Cultivation and identification of the transgenic alfalfa containing Echinococcus granulosus Eg95 gene
Yan-ju, YE ; Wen-gui, LI ; Hui, ZHOU
Chinese Journal of Endemiology 2009;28(3):283-287
Objective To cultivate and identify the transgenic affalfa containing Echinococcus granulosus Eg95 gene. Methods The alfalfa plants were transformed by co-cultivating alfalfa cotyledons via recombinant Agrobacterium tumefaciens LBA4404 harboring pBI-Eg95. The transgenic alfalfa explants were selected by kanamyein after calli formation, shoots and roots regeneration in the selective medium, the seedlings of transgenic plants were obtained which were finally transplanted into pots containing nutrient soil. After 2-3 months growth, the complete transgenic alfalfa plants containing Echinococcus granulosus Eg95 gene were obtained. To identify the transgenic alfalfa plants, the total DNA, RNA and leaf protein were extracted from fresh leaf tissue of the transgenic alfalfa plants and confirmed by PCR, RT-PCR, SDS-PAGE and Western blot assay. Results A specific band around 471 bp was amplified by PCR with total DNA, and the same band was obtained by RT-PCR with total RNA, which confirmed that the Eg95 gene was stably integrated into the transformed alfalfa genome. SDS-PAGE analysis showed that the relative molecular mass(Mr) of the expressed protein was about 16.5×103, consistent with the Eg95 protein, and the level of Eg95 expression was up to 0.06% of total soluble leaf protein by Bio-Rad Quantity one assay. Western blot verified the expressed protein was reactive with the sera of mice infected with Echinococcus granulosus. Conclusion The transgenic alfalfa plants containing Echinococcus granulosus Eg95 gene are successfully cultivated.
4.Study on the Differences of Virulence Genes and Molecular Typing in Campylobacter Jejuni Isolates from Poultry Products and Diarrhea Patients in Shenzhen
Bo LI ; Hui CHEN ; Changyan JU ; Pengwei HU
Journal of Modern Laboratory Medicine 2016;31(5):107-109,112
Objective To understand the differences of virulence genes and molecular typing in Campylobacterjejuni isolates from poultry products and diarrhea patients in Shenzhen.Methods According to specific primers,four virulence genes (cdtB,cadF,flaA,virB1 1 )of C.jejuni were detected by polymerase chain reaction (PCR).Molecular typing for C.jejuni strains was performed by Pulsed-field gel electrophoresis (PFGE).Results There were no differences of gene distribution (cdtB+,cadF+,virB1 1-)between isolates from poultry products and diarrhea patients.Two virulence genes of cdtB and cadF were found in all of ten C.jejuni strains lacking virB1 1 .The carriage rate of flaA in food-borne isolates (3/5 )was higher than those in patient isolates (2/5).In the PFGE map,the clustering analysis of C.jejuni strains showed that a total of 5 to 9 DNA bands were observed in ten strains through the digestion of Sam I.There was high homology (above 85%) between food-borne isolates and patients isolates,but the distribution of flaA in these highly homologous strains was differ-ent.Conclusion So far,C.jejuni strains with cdtB,cadF and flaA were present in Shenzhen,and showed high diversity and homology.This implies that the occurrence of diarrhea in patients with C.jejuni was associated with the contaminated poul-try products by this pathogen.Their findings can provide basic data and evidences about diarrheal disease caused by food-borne C.jejuni for the local region.
5.Clinical characteristics analysis of Chlamydia psittaci pneumonia detected by metagenomic next-generation sequencing
ZHAO Hui-jun ; OUYANG Xiao-li ; CHEN Ju-ping
China Tropical Medicine 2023;23(5):523-
Abstract: Objective To analyze the clinical characteristics of Chlamydia psittaci pneumonia and improve the diagnosis and treatment skills of clinicians on this disease. Methods The clinical data of thirty-nine Chlamydia psittaci pneumonia cases detected by metagenomic next-generation sequencing (mNGS) from September 2020 to January 2022 at the Affiliated Hospital of Southwest Medical University were retrospectively analyzed. Results There was a history of poultry exposure in 89.7%(35 cases) of the patients. The most common clinical manifestations were high fever (92.3%, 36), cough (76.9%,30), muscle soreness (48.7%,19), headache (38.5%,15), etc. Laboratory examinations showed 76.9% of patients had a normal leukocyte count, and 76.9% had decreased lymphocyte count, often accompanied by elevated C-reactive protein (100%), procalcitonin (97.4%), interleukin-6 (95.8%), interleukin-10 (95.8%), alanine aminotransferase (74.4%), and aspartate aminotransferase (84.6%). Univariate analysis indicated that there were statistically significant differences in the levels of aspartate transaminase, blood urea nitrogen, C-reactive protein, and procalcitonin between severe pneumonia patients and non-severe pneumonia patients(P<0.05). Multivariate logistic regression analysis showed that an elevated blood urea nitrogen (OR=4.899) had guiding significance for predicting the occurrence of severe pneumonia. Bronchoscopy examination showed no abnormalities in 53.6% of the patients. The imaging manifestations of pulmonary lesions were mainly lobar pneumonia (61.5%) and air bronchograms (94.9%). Therapeutically, it was sensitive to tetracyclines, macrocyclic lactones, and fluoroquinolones. A total of 84.6%(33 cases) of the patients were cured and discharged from the hospital at the end of the treatment. Conclusion Chlamydia psittaci pneumonia is a zoonotic disease that can be detected by mNGS. An elevated blood urea nitrogen level has guiding significance for predicting the occurrence of severe pneumonia. Empirically-selected regimens based on doxycycline are effective for the treatment of Chlamydia psittaci pneumonia.
6.Decoration reconstruction for finger defect in emergency
Ji-Hui JU ; Rui-Xing HOU ; Yue-Fei LIU ; Qiang ZHAO ; Lei LI ; Guang-Zhe JIN ;
Chinese Journal of Microsurgery 2006;0(06):-
Objective To discuss methods of decoration reconstruction for finger defect in emergency and to observe the elinical effects.Methods Of the 41 cases of finger injuries of different degrees,15 were repaired with part of the skin flaps of the big toenails or skin flaps of the second toenalis,8 were repaired with part of the skin flaps of the big toenails,7 were reeonstructed with the second tiptoes,11 were repaired with the abdominal skin flaps of the big toes or lateral flaps of the second toes.Results All the 41 fingers sur- vived.One skin flap of the big toe was somewhat swelling and a decorating operation was performed.The 4~18 months of follow-up visitation of the rest cases revealed good function and shapes.No obvious functional ab- norality was found in the donating feet.Conclusion Various kinds of decoration reeonstruetion for finger defects are available to recover the hand shape and function as much as possible.
7.Therapeutic Observation of Acupoint Sticking plus Hot Compress by Moxa-salt Bag for Constipation After Acute Cerebral Stroke
ju Hui LING ; hong Yu YAN ; na Li WANG
Shanghai Journal of Acupuncture and Moxibustion 2017;36(9):1061-1064
Objective To observe the clinical efficacy of acupoint sticking plus hot compress by moxa-salt bag in treating constipation after acute cerebral stroke.Method Eighty patients with constipation after cerebral stroke were randomly allocated to a treatment group and a control group by using random number table method according to registration order, 40 cases each. The control group was intervened by acupoint sticking at umbilicus with rawDa Huang (Rheum Officinale), and the treatment group was additionally given hot compress using moxa-salt bag. The treatment was given once a day, for consecutive 3 days in total. The first defecation time, stool shape and defecation sensation were observed in the two groups. The clinical efficacies were compared between the two groups.Result The total effective rate was 90.0% in the treatment group versus 72.5% in the control group, and the difference was statistically significant (P<0.05). After the treatment, there was statistically significant difference in comparing the first defecation time between the two groups (P<0.05). After the treatment, the incidence rate of soft stool was 85.0% and that of hard stool was 15.0% in the treatment group versus 40.0% and 60.0% in the control group, and the differences were statistically significant (P<0.05). The incidence rate of pleasant defecation sensation was 80.0% and that of difficult defecation was 20.0% in the treatment group versus 35.0% and 65.0% in the control group after the treatment, and the differences were statistically significant (P<0.05).Conclusion Acupoint sticking plus hot compress by moxa-salt bag is an effective method in treating constipation after acute cerebral stroke.
8.Study on the relationship between substance P and expressions of interleukin and interferon on colonic mucosa in patients with post-infective irritable bowel syndrome
Liang-Zhou WEI ; Hui JU ; Su-Mei DAI ; Lihua WANG ; Xishuang LIU ; Yanqing LI ;
Chinese Journal of Digestion 2001;0(09):-
Objective To investigate the effect of infection on the expression of substance P(SP) and Th1 cytokine on colonic mucosa in patients with irritable bowel syndrome(IBS).Methods Mucosa biopsy samples were obtained in descending colon and rectum from 77 patients(20 post-infective-IBS cases)and 30 controls by colonoscopy.The expressions of SP,interleukin(IL)-2,interferon(IFN)-?were determined by immunohistochemical staining.Results Compared with controls,expressions of SP,IL-2 and IFN-?were significantly increased in IBS patients with diarrhea-predominant(P<0.05),and expression of SP in post-infective-IBS patients with cytokines positive(IL-2 and IFN-?)were also signifi- cantly increased(P<0.05).There were no significant differences between non post-infective-IBS and controls.Conclusions Expression of SP was significantly increased in post-infective-IBS.SP upregulated the expression of Thl cytokines(IL-2 and IFN-?).Gastrointestinal infection may involve in the patho- genesis of IBS through neuroimmune mechanism.
9.Study on the Characteristics and Influential Factors of Microbial Rapid Detection Reagent by Bioluminescence
Ju-Mei ZHANG ; Qing-Ping WU ; Cheng-Si LI ; Hui-Qing WU ;
Microbiology 1992;0(03):-
In this assay, the reaction kinetics, optimum temperature, pH and various influential factors of ATP microbial rapid detection reagent by bioluminescence were studied. The results showed that it's enough for detection system to have 40 ~ 50?g/mL D-Luciferin. The light production decreased fastest in the first minute of reaction, then began to decay slowly. The optimal reaction temperature was 24℃~25℃and the optimal pH was pH 7.2 -7.4 in the reaction system. In addition, when stored at 4℃for 45h, the dissolved reagent solution could keep its 86% activity. When preserved at 25℃, the enzyme activity decreased less for 1h, and degraded gradually as time went by and only left 53. 5% of its activity after 6. 5h. While stored at 33℃, the enzyme activity decreased quickly with the time and only left 59. 1% after 1. 5h. The result indicated that storage temperature was a very important influential factor to the activity of reagent Meanwhile, different chemical substance such as acid, alkali, salt and surfactants inhibited the ATP bioluminescent reaction. When the concentration of NaCl reached 1. 5g/L, it could inhibit 52. 5% light production. Triton X-100, acid, and alkali also had some effects on the reaction, while CTAB, SDS and TCA would inhibit the bioluminescent reaction seriously.
10.Effect of different pressure CO2 pneumoperitoneum on function of liver and kidney in patients undergoing laparoscopic gastrectomy
Hui YE ; Shengbin WANG ; Xia JU ; Shenghong HU ; Siqi XU ; Yuanhai LI
The Journal of Clinical Anesthesiology 2015;(12):1168-1171
Objective To investigate the effect of different pressure CO2 pneumoperitoneum on function of liver and kidney in patients undergoing laparoscopic gastrectomy.Methods A total of sixty pa-tients,aged 40-65 years,scheduled for elective laparoscopic gastrectomy were randomly divided into three groups (n=20).The pressures of CO2 pneumoperitoneum were set at 6-8,9-11 and 12-14 mm Hg in group L,group M and group H respectively.The venous blood samples before pneumoperitoneum (T1 ),1 hour after pneumoperitoneum(T2 ),2 hour after pneumoperitoneum (T3 ),1 hour(T4 )and 24 hours(T5 )after stopping pneumoperitoneum were collected for determination of serum concentrations of NAG,Cys-C, ALT,AST,Cr,BUN and the amount of urine.pH,PaCO2 ,PaO2 were recorded at T1-T4 .Results ALT, AST,Cr and BUN were not different at T1-T5 in the three groups.Compared with group L,the concentra-tion of NAG and Cys-C at T2-T5 were significantly higher in the group M and H (P <0.05).Compared with group L,the amount of urine at T2-T5 was significantly lower in the group M and H (P <0.05 ), PaCO2 was significantly increased in the group H and M at T2-T4 (P <0.05).Conclusion The pressure of 6-8 mm Hg CO2 pneumoperitoneum can alleviate the damage of function of liver and kidney in patients un-dergoing laparoscopic gastrectomy.