2.Plasmid-mediated carbapenemase KPC-2 in a strain of Klebsieila pneumoniae
Xing-Guo ZHANG ; Xiao-Xing DU ; Rong ZHANG ; Ze-Qing WEI ; Yun-Song YU ; Ya-Gang CHEN ; Lan-Juan LI ;
Chinese Journal of Laboratory Medicine 2003;0(09):-
Objective To investigate the resistant mechanism of imipenem-resistant K. pneumoniae.Methods The minimal inhibitive concentrations (MICs) of the antimicrobial agents were determined by Etest.Isoelectric focusing electrophoresis (IEF),plasmid extraction,conjugation, transformation,PCR amplification,cloning and sequencing were carried out for analyzing the encoding gene of ?-1actamases.Results Three kinds of ?-1actamases were detected with pIs of 7.2,6.7,and 5.4.in a clinical strain of K.pneumoniae.These ?-1actamases were TEM-I (pI,5.4),SHV-12 (pI,8.2) and KPC-2 ( pI,6.7 ) confirmed by sequencing of the PCR products.Only one band of ?-1actamase with pI 6.7 was displayed in the transformant.A 1500 bp segment,which contained the KPC-2 gene confirmed by nucleotide sequence analysis,was cloned from a 60 000 bp plasmid of the transformant.Conclusion The strain of K.pneumoniae resistant to imipenem produces a plasmid-mediated carbapenemase KPC-2 which belongs to Bush group 2f,class A ?-1actamase.
3.Expression of nitric oxide and γ -aminobutyric acid in the retina of two kinds of amblyopia cats
Han-Min, WANG ; Ao, RONG ; Li-Juan, MO ; Qing-Song, LI ; Xing-Ru, ZHANG
International Eye Science 2016;16(11):2006-2009
AIM: To study the role of nitric oxide ( NO ) and γ-aminobutyric acid ( GABA) in the formation of amblyopia by establishing 2 different types of amblyopic models.METHODS:A total of 18 aged 3-week kittens were randomly divided into monocular deprivation, strabismus and normal groups. All types of amblyopia were developed in the experimental eyes that were detected by P-VEP 12wk later. The cats were killed and the immunocytochemistry staining method were applied to observe under the light microscope the changes of distribution and positive cells areas of NO and GABA across the amblyopic retinal, compared to that from the normal cats of identical age.
RESULTS: The P-VEP showed that the amplitude of wave P1 was lower (P<0. 05) and the P1 latent time was longer ( P<0. 05 ) in two types of amblyopic cats than those in the normal cats. Compared to the normal cats, the NO and GABA positive cells areas were obviously reduced ( P<0. 05 ) across the retina in the amblyopic cats. But no significant difference was found between two kinds of amblyopic cats.
CONCLUSION:The NO and GABA play an important role in the formation of amblyopia in the level of retinal.
4.Study of estradiol on treatment of preeclampsia in rat model
Xueyan WANG ; Qing XIONG ; Chao WANG ; Bing XIAO ; Shu ZHOU ; Rong ZHOU ; Aiyun XING
Chinese Journal of Obstetrics and Gynecology 2000;0(11):-
Objective To evaluate whether estradiol can inhibit and cure the inflammation of experimental preeclampsia in rats. Methods Experimental preeclampsia was induced in 14-day-pregnant rats by infusion of endotoxin (1.0 ?g/kg). Rats with normal pregnancy were infused with sodium chloride solution.A group of preeclampsia rats was injected with 17?-estradiol (17?-E_2, 1 mg?kg -1 ?d -1 ). Blood pressure, albuminuria,inflammation associated adhesion molecule CD_ 49d and tumor necrosis factor-?(TNF-?) were assessed. Results On pregnant day 19, for normal pregnancy group(group C) the blood pressure was (120.4?2.0)mm Hg (1 mm Hg=0.133 kPa),urinary protein (0.47?0.06)mg/24 hours;for experimental preeclampsia group(group A) blood pressure was (134.2?2.4) mm Hg,urinary protein(0.79?0.10)mg/24 hours; for experimental preeclampsia with 17?-E_2 treatment group (group B) blood pressure was(123.3?1.7)mm Hg,urinary protein (0.51?0.08)mg/24 hours. A significant increase of blood pressure and urinary albumin was observed in group A. CD_ 49d expression and TNF-? concentration were also increased. 17?-E_2 reduced the expression of CD_ 49d , concentration of TNF-?,blood pressure and albuminuria of experimental preeclampsia. However, the weight of fetuses in 17?-E_2 treatment group were less than that in other groups. Conclusion 17?-E_2 can improve the symptoms of experimental preeclampsia,but its effects on fetus need to be further studied.
6.Repair of acutely injured spinal cord through constructing tissue-engineered neural complex in adult rats.
Yu PU ; Qing-shan GUO ; Ai-min WANG ; Si-yu WU ; Shu-xing XING ; Zhong-rong ZHANG
Chinese Journal of Traumatology 2007;10(3):171-176
OBJECTIVETo construct tissue-engineered neural complex in vitro and study its effect in repairing acutely injured spinal cord in adult rats.
METHODSNeural stem cells were harvested from the spinal cord of embryo rats and propagated in vitro. Then the neural stem cells were seeded into polyglycolic acid scaffolds and co-cultured with extract of embryonic spinal cord in vitro. Immunofluorescence histochemistry and scanning electron microscope were used to observe the microstructure of this complex. Animal model of spine semi-transection was made and tissue-engineered neural complex was implanted by surgical intervention. Six weeks after transplantation, functional evaluation and histochemistry were applied to evaluate the functional recovery and anatomic reconstruction.
RESULTSThe tissue-engineered neural complex had a distinct structure, which contained neonatal neurons, oligodendrocytes and astrocytes. After tissue-engineered neural complex was implanted into the injured spinal cord, the cell components such as neurons, astrocytes and oligodendrocytes, could survive and keep on developing. The adult rats suffering from spinal cord injury got an obvious neurological recovery in motor skills.
CONCLUSIONSThe tissue-engineered neural complex appears to have therapeutic effects on the functional recovery and anatomic reconstruction of the adult rats with spinal cord injury.
Animals ; Female ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; surgery ; Stem Cell Transplantation ; methods ; Tissue Engineering ; methods
7.Histone deacetylase inhibitor promotes differentiation of embryonic stem cells into neural cells in adherent monoculture.
Xing YAO ; Jia-rong ZHANG ; Hua-rong HUANG ; Li-cheng DAI ; Qing-jun LIU ; Ming ZHANG
Chinese Medical Journal 2010;123(6):734-738
BACKGROUNDEmbryonic stem (ES) cells poss unlimited self-renewal capacity and the ability to differentiate into cell of all three germ layers in vitro. Induced differentiation of ES cells to neural lineage cells has great potential in basic study of neurogenesis and regeneration therapy of neurodegenerative diseases. Histone deacetylase (HDAC) inhibitors enhance histone acetylation so that globularly activate gene expression and may initiate multilineage differentiation. In this study, we aimed to develop a method to induce the differentiation of ES cells to neural cells combining HDAC inhibition and neural cell selection.
METHODSIn this study, we used HDAC inhibitor sodium butyrate (NaB) to induce the differentiation of mouse embryonic stem cells to neural cells through monolayer culture. After differentiation initiation by histone deacetylase inhibitor sodium butyrate, neural cells were induced and selected with a serum free culture system.
RESULTSHomogeneous neurons without glial cells demonstrated by molecular marker expression were differentiated with the method. The resultant neurons were excitable.
CONCLUSIONThe method combined differentiation induction effect of HDAC inhibitors and selective culture system to derive neural cells from ES cells, and implied the involvement of epigenetic regulation in neural differentiation.
Animals ; Butyrates ; pharmacology ; Cell Adhesion ; Cell Cycle ; drug effects ; Cell Differentiation ; drug effects ; Cells, Cultured ; Embryonic Stem Cells ; cytology ; drug effects ; Fibroblast Growth Factor 2 ; pharmacology ; Histone Deacetylase Inhibitors ; pharmacology ; Mice ; Neurons ; cytology ; physiology
8.Apoptosis in esophageal cancer cells induced by all-trans retinoic acid.
Tai-Ying LU ; Qing-Xia FAN ; Liu-Xing WANG ; Rui-Lin WANG ; Pei-Rong ZHAO ; Shi-Xin LU
Chinese Journal of Oncology 2007;29(11):822-825
OBJECTIVETo study the anti-tumor effects of all-trans retinoic acid (ATRA) and mechanisms of its action.
METHODSHuman esophageal carcinoma cell line EC9706 cells were treated with ATRA at different concentration. The proliferation inhibition was examined by MTT assay. Morphological examination, TUNEL method and flow cytometry were used to detect the apoptosis and changes of cell cycle. Immunohistochemical method was used to detect the expression of apoptosis-related genes caspase-3 and bcl-2. The semi-quantification of protein expression was analyzed by pathological image analysis.
RESULTSATRA inhibited the proliferation of EC9706 cells moderately. Apoptosis in EC9706 cells was induced by ATRA treatment. The morphology of EC9706 cells showed changes such as nuclear chromatin condensation and fragmentation. Sub-G1 peak was found by flow cytometry. The maximal apoptosis rate was 32.6%. The expression of caspase-3 gene was enhanced. The expression of bcl-2 gene was decreased. All these effects were presented in a dose-dependent and time-depend manner.
CONCLUSIONApoptosis is one of the key mechanisms of ATRA action on EC9706 cells.
Antineoplastic Agents ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Esophageal Neoplasms ; metabolism ; pathology ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Tretinoin ; administration & dosage ; pharmacology
9.Reproductive hormone levels and relevant parameters in middle-aged and older men in the urban area of Nanjing.
Xing-Rong QING ; Ling-Ling WANG ; Xu-Xin ZHAN ; Dun-Sheng MO ; Hong-Cai CAI ; Xue-Jun SHANG ; Shan-Chao ZHAO ; Yu-Feng HUANG
National Journal of Andrology 2014;20(4):347-353
OBJECTIVETo investigate the status of male reproductive health among middle-aged and older men in the urban area of Nanjing.
METHODSWe collected the laboratory results of 884 middle-aged and older men aged 55 - 89 years from the Xuanwu District of Nanjing present for routine physical examinations, including those of blood routine tests, liver and kidney function, blood glucose, blood lipid, and total prostate specific antigen (TPSA), as well as such reproductive hormone indexes as total serum testosterone (TT), free serum testosterone (fT), and sex hormone binding globulin (SHBG). We also obtained the above reproductive hormone indexes from 119 young and middle-aged men aged 20 - 39 years as controls.
RESULTSAging-related changes were found in the 50 percentiles of all the reproductive hormones and relevant parameters but those of TT and E2, with gradual increases in LH, FSH and SHBG and decreases in fT, TSI and fTI. Comparison of reproductive hormones and relevant parameters by Mann-Whitney U test did not show any statistically significant differences in the TT level between any two of the five age groups (20 - 39, 55 - 59, 60 - 69, 70 - 79, and > or = 80 yr) (P > 0.05) except between the control and > or = 80 yr groups and the 60 - 69 and > or = 80 yr groups (P < 0.05), nor in the E2 level between any two groups, nor in the levels of LH and FSH except between the 55 - 59 and 60 - 69 yr groups and the 70 - 79 and > or = 80 yr groups, and nor in the levels of fT and TSI except between the 55 - 59 and 60 - 69 yr groups. However, there were significant differences in the levels of SHBG and fTI between any two age groups. Spearman correlation analysis revealed that fT, TSI, and fTI were correlated negatively with aging and LH (P < 0.05, I r I > 0.5) but weakly positively with cholesterol, blood glucose and hemoglobin (P < 0.05, /r/ < 0.5), SHBG and LH positively with aging, SHBG weakly negatively with blood glucose and hemoglobin, LH weakly negatively with hemoglobin, and TT weakly negatively with aging but positively with hemoglobin.
CONCLUSIONThe levels of serum testosterone, particularly that of fT, declined with aging in middle-aged and older men in the urban area of Nanjing, which may contribute to abnormal lipid metabolism, low hemoglobin and high blood glucose.
Aged ; Aged, 80 and over ; Aging ; blood ; Blood Glucose ; analysis ; Case-Control Studies ; China ; Follicle Stimulating Hormone ; blood ; Humans ; Luteinizing Hormone ; blood ; Male ; Middle Aged ; Prostate-Specific Antigen ; blood ; Sex Hormone-Binding Globulin ; analysis ; Statistics, Nonparametric ; Testosterone ; blood
10.Lipopolysaccharide affects testicular histology and reproductive endocrine function in male rats.
Xu-Xin ZHAN ; Yu-An HU ; Xing-Rong QING ; Dun-Sheng MO ; Hong-Cai CAI ; Xue-Jun SHANG ; Qi ZHANG ; Yu-Feng HUANG
National Journal of Andrology 2014;20(4):304-308
OBJECTIVETo study the influence of lipopolysaccharide (LPS)-induced inflammation on the testicular histology and reproductive endocrine function in male rats and investigate the possible mechanism of inflammation affecting male fertility.
METHODSThirty-six male SD rats were randomly divided into a control group (A) and three LPS intervention groups (B, C, and D) to receive saline and LPS (5 mg/kg i. p, once), respectively. The animals in groups B, C, and D were killed by anesthesia at 12, 24, and 72 hours after treatment. Histopathological changes in the left testis of the rats were observed by HE staining and the levels of the reproductive hormones T, FSH, and LH in the serum were determined by ELISA.
RESULTSCompared with group B, group A showed clear structure of seminiferous tubules, orderly arrangement of spermatogenic cells, a slightly decreased number of sperm in some seminiferous tubular lumens, and shed spermatogenic cells in the rat testis tissue; group C exhibited thinner seminiferous epithelia, disordered structure of seminiferous tubules, irregular arrangement of spermatogenic cells, decreased number of mature sperm and obvious shedding of spermatogenic cells in seminiferous tubular lumens; group D manifested similar findings to those of group C, with even more shed spermatogenic cells that blocked the tubular lumens. The levels of serum T, LH, and FSH were (0.490 +/- 0.028) ng/ml, (6.290 +/- 0.515) ng/L, and (1.837 +/- 0.127) IU/L in group A, (0.460 +/- 0.024) ng/ml, (5.881 +/- 0.124) ng/L, and (1.707 +/- 0.098) IU/L in group B, (0.417 +/- 0.021) ng/ml, (5.123 +/- 0.271) ng/L, and (1.620 +/- 0.115) IU/L in group C, and (0.378 +/- 0.021) ng/ml, (4.504 +/- 0.279) ng/L and (1.562 +/- 0.216) IU/L in group D, all decreased in group B as compared with A (P > 0.05). The decreases of T and LH were extremely significant (P < 0.01) and that of FSH was significant in groups C and D (P < 0.05) in comparison with A.
CONCLUSIONLPS-induced inflammation affects the testicular tissue and reproductive endocrine function of male rats, resulting in decreased levels of serum T, LH, and FSH.
Animals ; Endocrine System ; drug effects ; physiology ; Fertility ; drug effects ; physiology ; Follicle Stimulating Hormone ; blood ; Humans ; Lipopolysaccharides ; toxicity ; Luteinizing Hormone ; blood ; Male ; Random Allocation ; Rats ; Reproduction ; Seminiferous Tubules ; drug effects ; pathology ; Spermatocytes ; drug effects ; Testis ; drug effects ; pathology ; Testosterone ; blood