Objective To clone lexA gene,express and purify the repressor LexA from Pseudomonas aeruginosa(PA),prepare the polyclonal antibody against PC-1 protein in rabbits,detect immunological activity of LexA protein.Methods The genomic DNA was extracted from the PAO1,the gene fragment encoding the mature LexA was amplied by PCR.It was linked the vector pET32a(+)and expressed in the E.coli BL21(DE3).The expressed protein was purified by two steps of Ni2+ chelate affinity chromatography and gel filtration chromatography respectively.The purified LexA protein immune the rabbits by injection and prepare the polyclonal antibody against PC-1 protein.The immunological activity of expressed and purified LexA protein was detected by ELISA,and Western blot.Results The expressed fused protein was found in insoluble form,accounted for 45% of the total bacteria protein.The final purity was 98.97%,which was determined by the HPLC.The expressed and purified LexA protein had satisfactory immunological activity.

OBJECTIVETo explore the mechanisms of Qianjing Decoction in the treatment of oligoasthenospermia (OAS).

METHODSWe randomly divided 100 SPF male rats into five groups of equal number: normal, model, Huangjingzanyu, levocarnitine, and Qiangjing. OAS models were established in the animals followed by intragastrical administration of normal saline, ornidazole, Huangjingzanyu Capsules (200 mg per kg body weight per day), levocarnitine (100 mg per kg body weight per day), and Qianjing Decoction (10 g per kg body weight per day), respectively, qd, for 4 successive weeks. Then, we detected the concentration and motility of the epididymal sperm, obtained the contents of superoxide dismutase (SOD), malonaldehyde (MDA), glutathione peroxidase (GSH-Px), lactate dehydrogenase (LDH), α-glucosidase, and fructose in the epididymis, and determined the mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and succinate dehydrogenase (SDH) in the epididymal tissue of the rats by real-time PCR.

RESULTSThe concentration and motility of the epididymal sperm in the model, Huangjingzanyu, levocarnitine, and Qianging groups were (35.34 ± 4.22) x 10(6)/ml and (40.04 ± 7.05)%, (48.12 ± 5.56) x 10(6)/ml and (62.46 ± 7.12)%, (47.14 ± 4.87) x 10(6)/ml and (63.23 ± 6.34)%, and (50.25 ± 5.08) x 10(6)/ml and (66.34 ± 7.58)%, respectively, all significantly lower than in the normal group ([53.05 ± 4.55] x 10(6)/ml and [70.20 ± 8.54]%) (P < 0.05), but remarkably higher in the Huangjingzanyu, levocarnitine, and Qiangjing groups than in the model rats (P < 0.05). Compared with the thinned epididymal lumen walls, decreased sperm count, and disorderly and loose arrangement of the lumens in the OAS models, the rats in the Huangjingzanyu, levocarnitine, and Qiangjing groups showed evidently thicker epididymal lumen walls, with the lumens full of sperm cells and arranged regularly and compactly, similar to those of the normal rats. The levels of SOD and GSH-Px were significantly lower but that of MDA markedly higher in the model rats ([84.12 ± 23.25], [10.56 ± 3.02], and [14.04 ± 2.06] nmol/mg) than in the normal group ([110.04 ± 19.56], [17.25 ± 3.56], and [8.87 ± 1.35] nmol/mg) (P < 0.05), while the former two indexes remarkably higher and the latter one significantly lower in the animals treated with Qiangjing Decoction ([120.56 ± 23.68], [16.34 ± 3.12], and [8.45 ± 1.56] nmol/mg), Huangjingzanyu Capsules ([115.34 ± 21.35], [15.23 ± 3.67], and [8.33 ± 1.54] nmol/mg), and levocarnitine ([116.67 ± 22.67], [15.35 ± 3.45], and [8.05 ± 1.78] nmol/mg) than in the models (P < 0.05). The levels of fructose, LDH and α-glucosidase were decreased markedly in the OAS models ([100.22 ± 12.12] mg/[ ml x g], [322 ± 46.13] U/[ ml x g], and [10.48 ± 2.33] U/[ml x g]) as compared with the normal rats ([128.12 ± 13.45] mg/[ml x g], [428 ± 35.12] U/[ml x g], and [15.34 ± 3.12] U/[ ml x g]) (P < 0.05), remarkably higher in the rats treated with Qiangjing ([130.23 ± 13.67] mg/[ml x g] [455 ± 51.50] U/[ml x g], and [18.56 ± 4.67] U/[ml x g]), Huangjingzanyu ([124.16 ± 14.02] mg/[ml x g], [ 419 ± 43.14] U/[ml x g], and [17.64 ± 4.08] U/[ml x g]), and levocarnitine ([123.34 ± 14.02] mg/[ml x g], [430 ± 31.80] U/ [ml x g], and [16.85 ± 5.55] U/[ml x g]) than in the models (P < 0.05). The Nrf2 mRNA expression was significantly reduced in the models as compared with the normal rats (P < 0.05) but remarkably increased in the Huangingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05). The SDH mRNA expression was significantly lower in the model than in the normal rats (P < 0.05) but markedly elevated in the Huangjingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05), remarkably higher in the Qiangjing than in the Huangjingzanyu group (P < 0.05).

CONCLUSIONOrnidazole induces OAS in rats, which is closely associated with excessive oxidation and energy metabolism dysfunction. Qiangjing Decoction can improve and even reverse ornidazole-induced OAS in rats as well as improve the ultrastructure of their testicular and epididymal tissues. Antioxidation and improvement of energy metabolism are probably the action mechanisms of Qiangjing Decoction in the treatment of OAS.

Animals ; Antioxidants ; Asthenozoospermia ; chemically induced ; drug therapy ; metabolism ; Carnitine ; pharmacology ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Energy Metabolism ; drug effects ; Epididymis ; metabolism ; Glutathione Peroxidase ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Oligospermia ; chemically induced ; drug therapy ; metabolism ; Ornidazole ; Random Allocation ; Rats ; Sperm Count ; Sperm Motility ; Spermatozoa ; drug effects ; physiology ; Succinate Dehydrogenase ; metabolism ; Superoxide Dismutase ; metabolism ; alpha-Glucosidases ; metabolism

Objective To observe the effect of Ti-6AL-4V particles on morphological and func- tional changes of osteoclast in vitro.Methods Mature osteoclasts separated from New Zealand Rabbits were cultured on glass slices and cortical bone slices.The experimental group was stimulated by Ti-6AL- 4V particles at concentration of 0.1 mg/ml.The cells were stained with TRAP at different culture time to observe the morphological variety.The bone resorption pits on bone slices were stained by toluidine blue and the resorption areas analyzed by computer image analysis software.Results Osteoclasts phagocy- tosed the particles,with irregular shapes,deeper TRAP stain and earlier apoptosis.Stimulation by Ti- 6AL-4V particles brought about larger area of bone absorption lacuna.Conclusion Osteoclasts have the ability to phagocytose Ti-6AL-4V particles,which leads to morphological and functional changes and enhances bone resorption.

OBJECTIVETo investigate the phenotype change of airway smooth muscle cell (SMC) in bronchial asthma and the effect of Zhichuan Capsule (ZCC) on it.

METHODSSixty male SD rats were randomized into 5 groups equally, the normal control group, the model group, and three treated groups treated with ZCC, Western medicine (WM) and ZCC + WM respectively. Excepting rats in the control group, they were all made into bronchial asthma model rats and received respective treatment. Histomorphology of the airway SMC was observed by electron microsope at the 4th week, and the expressions of smalpha-actin and sm-MHC were examined at the 2nd and the 4th week after provocation.

RESULTSAs compared with the control group, moyfilaments in SMC were lesser, secretive organelles were more and levels of smalpha-actin and sm-MHC expressions at the 4th week were lower in the model group (P < 0.05). While histomorphology of SMC in the WM group was as narmal, but some hypoxia changes were seen, such as the fracture of mitochondria cristae and deplation of matrix; histomorphology of SMC in the ZCC treated group and the ZCC + WM treated group were similar to that in the normal control group, and their expressions of smalpha-actin and sm-MHC were enhanced as compared to those in the model group (P < 0.05).

CONCLUSIONAirway SMC changes from contractile phenotype to synthetic phenotype in asthma rats, but the progress of change may be inhibited by ZCC.

Animals ; Anti-Asthmatic Agents ; therapeutic use ; Asthma ; drug therapy ; pathology ; Bronchi ; cytology ; pathology ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Myocytes, Smooth Muscle ; metabolism ; pathology ; Phenotype ; Phytotherapy ; Rats ; Rats, Sprague-Dawley

Chronic kidney disease(CKD)is a major public health problem worldwide. Understanding by doctors and laboratorians of the importance of reliable serum creatinine measurement in GFR estimation and of factors that may affect creatinine measurement is critical to ongoing public health efforts to improve the diagnosis and treatment of patients with CKD.We present an overview of the commonly used methods,their performances and limitations and the required performance criteria for the measurement of serum creatinine.Available resources for standardization of serum creatinine measurements and recommendations for creatinine measurement and GFR estimations are introduced.

Objective To investigate the changes of plasma somatostatin(SS) and its correlation with cellular immune function in neonates with hypoxic-ischemic encephalopathy(HIE).Methods Fifty cases of HIE were collected to detect the SS and T lymphocyte subsets,IL-2,sIL-2R as well as IL-6 levels by radioimmunoassay,APAAP and doule antibody sandwith ELISA methods.Results The SS and sIL-2R levels in patients with HIE were significantly higher(P

OBJECTIVETo investigate the effects of epidermal growth factor (EGF) on estrogen receptor (ER) and androgen receptor (AR) in mouse prostate cells and explore the putative role of EGF in prostatic hyperplasia.

METHODSSixty male Kunming mice were randomly divided into two EGF groups and one control group (n=20) and subjected to subcutaneous injection of 1 and 2 microg/day EGF and distilled water, respectively, for 28 consecutive days. The cellular expression of ER and AR in the prostate of mice in different groups was evaluated by flow cytometry.

RESULTSCompared with the control group, the positivity rate of ER and its expression level were significantly increased in the mouse prostate after EGF treatment (P<0.01), and the ER expression level was significantly higher in mouse with 2 microg/day EGF treatment than in those treated with 2 microg/day EGF (P<0.01). AR positivity rate and expression level also increased significantly in comparison with the control group (P<0.05), but no significant variation was found between 1 microg/day and 2 microg/day EGF groups.

CONCLUSIONEGF can increase the cellular expression of ER and AR in mice prostate and may play a role in the pathogenesis of prostatic hyperplasia.

Animals ; Epidermal Growth Factor ; administration & dosage ; pharmacology ; Flow Cytometry ; Injections, Subcutaneous ; Male ; Mice ; Prostate ; cytology ; metabolism ; Prostatic Hyperplasia ; metabolism ; pathology ; Random Allocation ; Receptors, Androgen ; biosynthesis ; Receptors, Estrogen ; biosynthesis

Objective To summarize the experience of totally endoscopic thyroidectomy (TET) in treating differentiated thyroid carcinoma and to evaluate its feasibility, safety and the therapeutic result.Methods The clinical data of 25 cases of thyroid carcinoma treated with TET via the areola of breast or by axillary approach from November 2004 to July 2009 in our hospital were retrospectively analyzed.Result TET procedures were successfully performed in all 25 cases. Postoperative pathology confirmed that 23 cases were of papillary cancer and the other 2 were follicle cancer. Lobectomy and subtotal thyroidectomy were performed in 11and 14 cases respectively. In 7 cases additional lymph node dissection was performed.All the cases were followed-up, ranging from 5 to 58 months, with a median of 28 months. No recurrence was found during this period. There was no morbidity nor major complications and patients were all satisfied with the cosmetic effect. Conclusion Totally endoscopic thyroidectomy (TET) is curative, safe and cosmetic therapy for differentiated thyroid carcinoma.

Objective To observe the relationship between apoptosis of K562 cell induced by iron-deprivation and activation of Caspase-3.Methods K562 cells were treated with desferrioxamine(DFO) in different dosages were collected at different time points.K562 cells were labelled with Annexin V/PI,and then the rate of apoptosis was measured by flow cytometry;The activation of Caspase-3 were detected by colorimetric method with pAN labelled substrate;The active protein of Caspase-3 were analyzed by Western blot.Results When K562 cells were treated with different concentrations of DFO,the apoptosis rate and the activity of Caspase-3 increases gradually.When K562 cells were incubated with DFO(50 ?mol/L and 100 ?mol/L) 24 h later,the enzymatic activity of Caspase-3 increases dramatically more than that of control group,and the difference was significantly(P0.05).All those effect above can be counteracted by equal mole concentration of FeCl_3.Conclusion Iron-deprivation maybe induce the apoptosis of K562 cell by chelating intracellular iron and activing Caspase-3.

Adult ; Female ; Helicobacter Infections ; complications ; Helicobacter pylori ; Humans ; Hyperemesis Gravidarum ; etiology ; Pregnancy