2.Regeneration of cochlear hair cells of guinea pigs following gentamicin ototoxicity
Yueqiu NI ; Hao TANG ; Cheng CUI
Chinese Journal of Tissue Engineering Research 2006;10(30):177-179
BACKGROUND: Formerly, it was thought that the damaged hair cells could not have the repair ability. Recent studies demonstrate that mammal vestibule hair cells also possess certain repair ability after being destroyed.Then, whether mammalia animal cochlea hair cells possess regenerative ability after being destroyed is disputed.OBJECTIVE: To observe cochlear hair cells condition and threshold value change of auditory brainstem response (ABR) at different time following gentamicin ototoxicity by using scanning electron microscope (SEM) technique combined with ABR test, so as to investigate whether cochlear hair cells of mammals can be regenerated after being injured.DESIGN: A randomized and controlled animal experiment.SETTING: Department of Physiology, Shenyang Medical College.MATERIALS: This experiment was carried out at the Hearing Research Room of China Medical University from November 2001 to May 2002. Totally 60 healthy adult white Guinea pigs, with red eyes and sensitive auricle reflex, of clean degree, were used and randomly divided into gentamicin group and normal control group with 30 guinea pigs in each one.METHODS: 100 mg/kg gentamicin was intraperitoneally daily injected into the guinea pigs, serving as gentamicin group. Same volume of normal saline (2.5 mL/kg) was intraperitoneally daily injected into the guinea pigs,serving as normal control group. All the guinea pigs were given medication for 10 successive days. Threshold value of ABR was detected respectively pre-operatively and at the 1st, 3rd, 30th days postoperatively; after withdrawal and execution, scanning electron microscope was used to observe cochlear hair cells of guinea pigs in each group.MAIN OUTCOME MEASURES: ① Threshold value of ABR. ②Cochlear hair cell change of guinea pigs at different time following gentamicin ototoxicity.RESULTS: All the 60 experimental animals entered the stage of result analysis with no loss in the midway. ①At 1,3 and 30 days after withdrawal of gentamicin, threshold value of ABR was significantly higher as compared with normal control group, with significant difference [(38.00±3.75), (2.22 ±3.63) dB nHL,t=30.651, P < 0.001];[(39.09±4.22), (2.50±3.54) dB nHL, t=29.708, P < 0.001];[(14.50±3.69), (1.50±2.42) dB nHL,t =13.175, P < 0.001]. Threshold value of ABR recovered obviously on day 30, but did not reach the normal level. ②On the first day after withdrawal of gentamicin , stereocilium of hair cells in second turn of cochlea of guinea pigs presented fusion, distortion, lodging, loss or incompetence and other pathological changes , especially severe in the third turn , also cystic form protrutions appeared outside the stereocilium of inner hair cell; On day 3 after withdrawal of gentamicin, stereocilium of outer hair cells in the second turn of cochlea of guinea pigs still presented fusion, loss, lodging and other pathological changes. Stereocilium of inner hair cell still showed lodging,but the outside cystic form protrutions decreased; On day 30 after withdrawal of gentamicin, stereocilium of outer hair cells in the second turn of cochlea presented fusion, loss , lodging and other pathological change ,which were obviously weaker than those on the 1st day and 3rd day after withdrawal of gentamicin , and at the same time , new born stereocilium appeared in the third turn of cochlea.CONCLUSION: Cochlear hair cell morphology recovery appears in those which survive for 30 days after cochlear hair cells of guinea pigs are damaged following gentamicin ototoxicity, and threshold value of ABR also recovers to some extent, suggesting that cochlear hair cells possess regenerative and repair ability following gentamicin ototoxicity. Hair cells after gentamicin-induced cochlear damage possess regenerative ability.
3.Cdc25B overexpression reverses two-cell block in mouse embryos.
Cheng CUI ; Hao TANG ; Bingzhi YU
Chinese Journal of Applied Physiology 2008;24(1):99-103
AIMTo explore the effect of Cdc25B overexpression on the development of mouse two-cell embryos.
METHODSThe pBSK-Cdc25B was in vitro transcribed into 5'-capped mRNA for microinjection by using mMESSAGE mMACHINE kit. The Cdc25B mRNA was microinjected into mouse embryos at two-cell stage in order to observe the embryonic development and cleavage rate. Using protein kinase activity assay and Western blot to detect the MPF activity as well as the phosphorylation status of Cdc2-Tyr15 in Cdc25B overexpression group respectively.
RESULTSThe mouse embryos with Cdc25B overexpression developed to the four-cell stage 48 h after the hCG injection with the percentage of cleavage over 40% compared with the embryos in control groups which still remained at the two-cell stage. Moreover, MPF activity increased significantly after Cdc25B mRNA injection. The phosphorylation status of Cdc2-Tyr15 was coincident with MPF activity.
CONCLUSIONThe results indicate that Cdc25B overexpression in early mouse two-cell embryos reverses two-cell block and promotes their development into four-cell stage by activating MPF.
Animals ; Cell Cycle ; Cell Division ; Embryo, Mammalian ; cytology ; Embryonic Development ; physiology ; Female ; Male ; Maturation-Promoting Factor ; metabolism ; Mice ; Microinjections ; Mitosis ; RNA, Messenger ; metabolism ; Zygote ; growth & development ; metabolism ; cdc25 Phosphatases ; physiology
4.Magnetic resonance cholangiopancreatography to analyze the risk factor before laparoscopic cholecystectomy
Hao TANG ; Chuxiao SHAO ; Yonghong XU ; Xiaoxi FAN ; Feng CHENG ; Hua DONG
Chinese Journal of Digestive Endoscopy 2009;26(2):76-78
Objective To assess the diagnostic value of MRCP before LC.Methods 944 cases with chronic calculous cholecystitis underwent MRCP before LC from June 2004 to June 2007 in our department.incidence rate of cholecvstolithiasis together with common bile duct stones and incidence rate of anatomic abnormity of bile duct were collected.Results The incidence rate of cholecvstolithiasis together with common bile duct stones were 8.1%(77/944),and the oecurence ofACBDS were 1.2%(11/944).The incidence rate of anatomic abnormity of bile duct were 3.7%(35/944).ConclusionMRCP can not only offer a excellent diagnostic value of ACBDS and anatomic abnormity of bile duct,but also reduce the occurrence of CBDS remainder and iatrogenic bile duct iniuries.
5.Simultaneous Determination of Four Components in Huangqi Shengmai Decoction by HPLC-DAD
Cheng LU ; Shenghui ZHOU ; Wei LI ; Xiuli ZHENG ; Qingjian GAO ; Hao TANG
Chinese Journal of Information on Traditional Chinese Medicine 2015;(7):96-99
Objective To establish a method for simultaneous determination of contents of schisandrin B, schisantherin A, lobetyolin and ruscogenin in Huangqi Shengmai Decoction with HPLC method. Methods The chromatographic column was an Agilent Eclipse C18 column (4.6 mm × 250 mm, 5 μm) that was maintained at a constant temperature of 30 ℃. The mobile phase was 0.1% acetic acid solution-acetonitrile, with gradient elution. The flow rate was 1.0 mL/min. The detection was at wavelength of 280 nm. Results The standard curves of schisandrin B, schisantherin A, lobetyolin and ruscogenin had good linearity in the ranges of 0.862 5-21.562 5μg (r 2=0.999 1), 0.737 5-18.437 5μg (r 2=0.999 4), 0.095 2-2.380 0 μg (r 2=0.999 6), and 0.810 0-20.250 0 μg (r 2=0.999 0), respectively. The average recoveries of the four components were 98.06%(RSD=1.64%), 99.61%(RSD=2.72%), 97.98%(RSD=1.45%), and 99.30%(RSD=1.25%), respectively. Conclusion This method is accurate, rapid and specific, achieving the stable results with high reproducibility.
6.The absorption and metabolism differences of intestine and liver for multicomponent licorice water extract
Lei ZHANG ; Yanli PAN ; Yang LIU ; Mingmin TANG ; Li WEI ; Chengbo HOU ; Xiao CHENG ; Hao WANG
International Journal of Traditional Chinese Medicine 2015;(7):636-640
Objective This paper was to study the absorption and metabolism differences of intestine and liver for multicomponent licorice.Methods The components were identified with the UPLC-MS/MS. In situ closed-loop method was used to carry out the comparative experiments of absorption and metabolism differences between intestine and liver.Results 13 components were identified by UPLC-MS/MS. The absorption and metabolism results indicated some components in licorice water extract could be absorbed into blood and metabolism happened during this process. 14 metabolites were detected in the plasma sample. The hepatic metabolism results indicated many components could experience complex metabolism and more metabolites could be generated.Conclusions Liver was the major metabolism organ for licorice water extract and some components could be metabolized along with the absorption process in intestine. The absorption and metabolism differences between intestine and liver were significant.
8.Study on the B cell linear epitopes of rabies virus CVS-11 nucleoprotein.
Xin-Jun LV ; Xin-Xin SHEN ; Peng-Cheng YU ; Hao LI ; Li-Hua WANG ; Qing TANG ; Guo-Dong LIANG
Chinese Journal of Virology 2014;30(3):253-256
To study the B cell linear epitopes of rabies virus CVS-11 nucleoprotein, peptides were synthesized according to the amino acid sequences of B cell linear epitopes. Linear epitopes predicted by bioinformatics analysis were evaluated with immunological techniques. Indirect enzyme-linked immunosorbent assay showed that titers of antibodies to peptides (355-369 and 385-400 residues of rabies virus CVS-11 nucleoprotein) were above 1:12 800 in mouse sera. The antibodies recognized denatured rabies virus CVS-11 nucleoprotein in Western blot analysis. Purified anti-peptide antibodies recognized natural rabies virus CVS-11 nucleoprotein in BHK-21 cells in indirect fluorescent antibody test. The 355-369 and 385-400 residues of rabies virus CVS-11 nucleoprotein were validated as B cell linear epitopes.
Amino Acid Sequence
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Animals
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Antibodies, Viral
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immunology
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Epitope Mapping
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Epitopes, B-Lymphocyte
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chemistry
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genetics
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immunology
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Female
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Humans
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Male
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Mice
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Mice, Inbred BALB C
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Molecular Sequence Data
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Nucleoproteins
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chemistry
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genetics
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immunology
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Rabies
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immunology
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virology
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Rabies virus
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chemistry
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genetics
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immunology
9.Effects of injectio Salvia miltiorrhiza on gentamicin-induced free radical formation in guinea pig cochlea.
Ai-Mei WANG ; Hao TANG ; Jing SHEN ; Cheng CUI
Chinese Journal of Applied Physiology 2004;20(4):406-409
AIMTo investigate the effects of injectio Salvia Miltiorrhiza (SM) on gentamicin (GM)-induced free radical formation in guinea pig cochlea, and to explore possible mechanisms on GM-induced ototoxicity.
METHODSBiochemical assays of superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in guinea pig cochlea, combined with auditory brainstem response (ABR) measurement and transmission electron microscopic observation were used in this investigation.
RESULTSSOD activity was significantly declined while MDA content was distinctly increased in cochlear tissues after GM injection (P < 0.01). Moreover, they were well correlated with auditory function damage (|r| > 0.7). Co-treatment with SM evidently enhanced SOD activity and decreased MDA content (P < 0.01, P < 0.05). Furthermore, auditory function was markedly ameliorated. Morphological changes of cochlea were consistent with those of hearing function.
CONCLUSIONLipid peroxidation elicited by free radical was involved in GM-induced cochleotoxicity. SM might enhance SOD activity and prevent lipid peroxidation. As the result it might alleviate GM ototoxicity, and improve auditory function.
Animals ; Cochlea ; drug effects ; metabolism ; Free Radicals ; metabolism ; Gentamicins ; toxicity ; Guinea Pigs ; Lipid Peroxidation ; Malondialdehyde ; analysis ; Salvia miltiorrhiza ; Superoxide Dismutase ; metabolism
10.Surveillance on Antibiotic Susceptibility of Neisseria Gonorrhoeae from 1988 to 2002 in Shanghai
Weiming GU ; Yang YANG ; Lei WU ; Zhiqin GAO ; Chuguang ZHANG ; Quangui TANG ; Hao ZHANG ; Weizhong HU ; Ying CHENG ; Haishan WANG
Chinese Journal of Dermatology 1994;0(06):-
Objective To study the susceptibility of Neisseria gonorrhoeae to antibiotic agents from 1988 to 2002 in Shanghai. Methods The clinical isolates from patients with gonorrhea were collected and tested for their susceptibility to five antibiotics. Agarose-dilution-method was used to detect minimal inhibitory concentration (MIC) of anti-microbial agents including penicillin, tetracycline, spectinomycin, ciprofloxacin and ceftriaxone, and penicillinase producing Neisseria gonorrhoeae (PPNG) were tested with acidometric method. Results Susceptible strains to penicillin decreased from 11.28% in 1988 to 0 in 2002, MIC50 and MIC90 increased 8 and 4 times, respectively, the resistant rate and proportion of PPNG were 94.29% and 50.95%, respectively in 2002. The strains of high resistance to tetracycline increased from 0 in 1995 to 20.95% in 2002. The susceptible strains to ceftriaxone decreased from 100% in 1995 to 23.80% in 2002. The susceptibility to ciprofloxacin decreased significantly and resistant rate reached 99.05% in 2002. However, these strains were kept highly susceptible to spectionmycin. Concerning the multi-drug resistance, we found that the strains resistant to penicillin, ciprofloxacin and tetracycline simultaneously increased from 20.87% in 2001 to 23.30% in 2002, those resistant to both penicillin and ciprofloxacin reached to 70% in the past 2 years. Conclusions In Shanghai the resistant rates of Neisseria gonorrhoeae to antibiotics increased year by year in the past 15 years. The study indicates that spectinomycin and ceftriaxone should be the first choice for the treatment of gonorrhea at present and new sensitive antibiotic should be developed for the treatment of gonorrhea.