The DNA fragment sized 2 139bp, the same Sequence with AtKup1 gene from Arabidopsis thalianna was used as the templates for DNA family shuffling. The shuffeld AtKup1 gene library was expressed in the mutant of 5. cerevisae in which potassium transporter gene TRK1 and TRK2 were knocked out by homologous recombination. Then the screening was carried out in the low potassium media containing 5. 0 mmol/L KC1 and no histidine in it. it was found that both of diverse and wild AtKup1 gene can rescues the trk1△trk2△yeast mutant strain in low [ K + ] medium. The growth of 2 clones yeast containing diverse AtKup1 were beter than that of AtKup1 wild gene transformant. The sequencig results of the shuffeld AtKup1 showed that there were 2 nucleotide changed, which resulted in 2 amino acid variations in it compared with the original AtKup1. The potassium uptaking capacity of shuffled AtKup1 gene increased significantly when it was transformed into tobacco.

Potassium ion channels are a complex of protein molded in the cell membrane lipids. Its expression is strong in normal prostatic epithelia and weak in different degrees in prostatic cancer epithelia, but not clearly known in chronic prostatitis epithelia. Drugs affecting potassium ion channels could provide a new direction and some new ideas for the treatment of prostatic diseases.
Animals ; Humans ; Male ; Mice ; Potassium Channel Blockers ; pharmacology ; therapeutic use ; Potassium Channels ; classification ; drug effects ; physiology ; Prostatic Diseases ; drug therapy ; physiopathology

·AIM:To study the effects of timolol and latanoprost on optic disc parameters and retinal nerve fiber layer thickness of high intraocular pressure type POAG patients, and to provide guidance for clinical research.·METHODS: Totally 240 patients ( 336 eyes ) with high intraocular pressure type POAG in our hospital from November 2013 to November 2015 were randomly divided into control group and observation group, and each with a total of 120 cases (170 and 166 eyes, respectively). The patients in control group only received latanoprost treatment, while the observation group was treated with timolol and latanoprost treatment. After 3mo of treatment, we observed the therapeutic effects, and measured the optic disc parameters and retinal nerve fiber layer thickness. The incidence of adverse reactions was observed in the following 1a.·RESULTS: The patients of observation group and the control group after treatment showed the improvement rates of 97. 6% and 80. 6%, respectively, and the observation group was significantly better than the control group, the two groups showed statistically significant (P<0. 05). The eye rim area, rim volume, rim volume and diameter of two groups after treatment were higher than before ( P < 0. 05 ) , while those of the observation group was significantly higher than control group ( P<0. 05 ) . Two groups of patients with vertical cup to disc diameter decreased obviously ( P<0. 05 ) , the observation group was significantly lower than the control group, the difference was statistically significant ( P<0. 05 ) . Between the two groups of patients, during the period, incidences of dry eyes, corneal and conjunctival hyperemia, infiltration blurred vision and other complications had no significant difference ( P>0. 05), and the 1a follow-up of patients showed no other adverse reactions.·CONCLUSION: The use of timolol and latanoprost therapy for high intraocular pressure type POAG patients showed that the patients improve obviously, the visual acuity has more improved and the parameters of optic disc and retinal nerve fiber layer thickness are also improved.

OBJECTIVETo investigate the regulatory effect of potassium channel blocker (tetraethylammonium [TEA], aminopyridine [4-AP], glibenclamide [Glib]) on the proliferation of SD rat prostatic epithelial cells in vitro.

METHODSThe primary culture was prepared by collagenase dissociation of minced prostatic tissues. Cells were cultured in serum-free prostate epithelial cell growth media and identified by immunocytochemical studies. TEA and 4-AP at the concentration of 1, 5 and 10 mmol/L and Glib at the concentration of 10, 50 and 100 mol/L were added, and after 24, 48 and 72 hours of culturing, a cell column diagram was drawn and the cell number counted. The post-passage cell growth was observed by MTT assay and Hoechst33258 nucleus staining.

RESULTSThe cultured cells showed the typical morphological features of epithelia, with positive stain. MTT assay and Hoechst33258 staining showed that TEA, 4-AP and Glib at the increasing concentration effected different degrees of proliferation of prostatic epithelial cells after 24, 48 and 72 h (P < 0.01).

CONCLUSIONThe potassium channel blocker is a direct physiological regulator of the proliferation of SD rat prostatic epithelial cells.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Epithelial Cells ; drug effects ; Male ; Potassium Channel Blockers ; pharmacology ; Prostate ; cytology ; drug effects ; Rats ; Rats, Sprague-Dawley

The purpose of this study is to investigate the function of a novel potassium transporter gene (NrHAK1) isolated from Nicotiana rustica roots using yeast complement and real-time PCR technique. The complementary DNA (cDNA) of NrHAK1, 2 488 bp long, contains an open reading frame (ORF) of 2 334 bp encoding a protein of 777 amino acids (87.6 kDa) with 12 predicted transmembrane domains. The NrHAK1 protein shows a high sequence similarity to those of high-affinity potassium transporters in Mesembryanthemum, Phytolacca acinosa, Arabidopsis thaliana, and so on. We found that the NrHAK1 gene could complement the yeast-mutant defect in K+ uptake. Among several tissues surveyed, the expression level of NrHAK1 was most abundant in the root tip and was up-regulated when exposed to potassium starvation. Moreover, the transcript accumulation was significantly reduced by adding 5 mmol/L NH4+ to the solution. These results suggest that NrHAK1 plays an important role in potassium absorption in N. rustica.
Cation Transport Proteins ; chemistry ; genetics ; physiology ; Plant Proteins ; chemistry ; genetics ; physiology ; Potassium ; metabolism ; Quaternary Ammonium Compounds ; pharmacology ; Sodium ; pharmacology ; Tobacco ; genetics ; metabolism

Objective To study the expression of CD269 and CD317 antigens in bone marrow cells of patients with multiple myeloma (MM),analyze its correlation with the laboratory indexes reflecting the progression of MM and evaluate its value in clinical diagnosis and treatment.Methods 63 newly diagnosed MM patients were selected as the study group by a casecontrol study.The expression rate of CD269 and CD317 in bone marrow blood of 35 patients with iron deficiency anemia and other antigens in bone marrow blood of 63 patients with MM were detected by flow cytometry.The levels of serum hemoglo bin (Hb),serumβ2-MG(β2-MG) and lactatedehydrogenase (LDH) in patients with MM were dctectcd,and the levels of CD269 and CD317 were analyzed statistically.Results The positive rates of CD269 in the study group and control group were (86.6±2.35)％ vs (4.33±l.69)％,rcspectivcly (t =4.256,P＜0.05)).The positive rate of CD317 was (71.42+ 0.62)％ vs (8.32+ 3.89)％,the difference was statistically significant (t=3.102,P＜0.05).In other expression,the expression level of CD269 and CD317 in CD56 positive group was significantly higher than that of negative group (t=4.032,P＜0.05),while the expression of CD117 the level of positive group was significantly lower than that of the negative group (t 2.832,P＜0.05),CD19,CD20 expression was not statistically significant difference between the two groups (P＞ 0.05).The levels of CD269 and CD317 in patients with MM were positively correlated with the level of CD56 expression (r =0.392,P＜0.05),and negatively correlated with the level of CD117 expression (r=-0.210,P＜0.05).The levels of CD269 and CD317 in patients with MM were significantly lower than those in the negative group (t=3.012,P＜0.05) and the levels of serum LDH in the positive group were lower than those in the negative group (t=2.024,P＜0.05).There was a negative correlation between Hb content (r=-0.212,P＜0.05) and negatively correlated with serum β2-MG (r=-0.312,P＜0.05).Conclusion The high expression of CD269 and CD317 in bone marrow cells in MM patients is related to the increase of CD56 and decrease of CD117 in patients with MM.

OBJECTIVETo compare the bidirectional effect of rhubarb total anthraquinone (TA) and total tannins (TT) on rats' liver.

METHODSOne hundred rats were randomly divided into 10 groups, i.e., the blank group, the model group, the blank + high dose TA group, the blank +low dose TA group, the blank + high dose TT group, the blank + low dose TT group, the model + high dose TA group, the model + low dose TA group, the model +high dose TT group, and the model + low dose TT group, 10 in each group. The carbon tetrachloride (CCI4) was used to prepare the acute liver injury rat model. TA and TT of rhubarb (at 5.40 g crude drugs/kg and 14.69 g crude drugs/kg) were intragastrically administrated to rats in all groups except the blank group and the model group, once daily for 6 successive days.The general state of rats, biochemical indices such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), laminin (LN), hyaluronic acid (HA), transforming growth factor beta1 (TGF-beta1), as well pathological results of rat liver tissues. Finally the protection laws of TA and TT for rats' liver were analyzed using factor analysis.

RESULTSCompared with the blank control group, all biochemical indices increased in the blank group (P < 0.05, P < 0.01). HA also increased in the blank + high dose TA group; AST, ALT, and HA also increased in the blank +high dose TT group (P < 0.05). Compared with the model group, AST, ALT, ALP, HA, and TGF-beta1 significantly decreased in the model + low dose TA group, the model + high dose TA group, the model + low dose TT group (P < 0.05, P < 0.01). Serum AST, ALT, and ALP also decreased in the model + high dose TT group (P < 0.05, P < 0.01). Pathological results showed that mild swollen liver cells in the model + high dose TA group. Fatty degeneration and fragmental necrosis around the central veins occurred in the blank + high dose TA group. The pathological injury was inproved in the model +low dose TA group. Two common factors, liver fibrosis and liver cell injury, were extracted by using factor analysis. TA showed stronger improvement of the two common factors than TT.

CONCLUSIONSRhubarb TA and TT showed protective and harmful effects on rats' liver. At an equivalent dosage, TA had better liver protection than TT. High dose TT played a role in liver injury to some extent.

Animals ; Anthraquinones ; adverse effects ; pharmacology ; Carbon Tetrachloride ; toxicity ; Chemical and Drug Induced Liver Injury ; drug therapy ; pathology ; Dose-Response Relationship, Drug ; Female ; Liver ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Rheum ; chemistry ; Tannins ; adverse effects ; pharmacology

Objective To explore the differential expression of CD269 and CD317 in patients with multiple myeloma(MM). Methods Newly diagnosed samles from patients of MM(20 cases)and iron deficiency anemia(20 cases),40 cases in total (from 06/2015 to 08/2013,the Department of Hematology,Central Hospital of Zhuzhou City)were collected.Real-time quantitative PCR(RQ-PCR)tests were used to detect the relative expression of CD269 and CD317 in bone marrow sam-ples,and the results were statistically analyzed with clinical features.Results The relative expression levels of CD269 and CD317 in patients with multiple myeloma(4.418±4.568,4.327±2.876)were significantly higher than those in the control group(0.600±0.838,1.033±1.335),the difference was statistically significant(t=3.676,4.646,all P<0.05)respective-ly,while not related with the gender,age(P>0.05).There was no correlation between the expression of CD269 and CD317 (r=0.041,P=0.864),but positively correlated with the ratio of myeloma cells(r=0.495,P=0.026;r=0.533,P=0.016).Conclusion CD269 and CD317 were highly expressed in patients with multiple myeloma and may be involved in the pathogenesis of multiple myeloma.

OBJECTIVETo explore the feasibility of transfecting DHFR (human double-mutant dihydrofolate reductase) gene into mouse bone marrow cells and the effect of resistance to high dose MTX chemotherapy.

METHODSAfter DHFR gene was transfected into mouse bone marrow cells with retroviral vector, the cells were treated with methotrexate (MTX) and then CFU-GM (granulocyte-macrophage colony-forming unit) assay was performed. Peripheral blood leucocytes and platelets, body weight and survival rate were observed. After treatment with high dose MTX, the expression of drug resistance gene was checked by RT-PCR in the transfected bone marrow cells.

RESULTSSFG-F/S-NeoR gene-transfected mice bone marrow cells yielded drug-resistance colonies to MTX (donor mice: 15.8%, recipient mice: 18.0%, control: 0) The peripheral blood leucocytes and platelets, body weight recovered gradually and the survival rate was 83.3% at the 40th day, while 0 in controls in gene transfected mice after large dose MTX treatment. RT-PCR of transgenic mouse marrow cells showed the band of F/S gene (400 bp).

CONCLUSIONDHFR gene can not only be integrated and expressed in bone marrow cells but also improve their drug-resistence to MTX.

Animals ; Antimetabolites, Antineoplastic ; pharmacology ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Bone Marrow Transplantation ; Cells, Cultured ; Drug Resistance, Neoplasm ; genetics ; Erythrocyte Count ; Genetic Vectors ; Leukocyte Count ; Male ; Methotrexate ; pharmacology ; Mice ; Mice, Inbred BALB C ; Mutation ; Retroviridae ; genetics ; Survival Analysis ; Tetrahydrofolate Dehydrogenase ; genetics ; metabolism ; Transfection

OBJECTIVETo analyses the clinical application of thoracodorsal artery perforator flaps (TAP).

METHODSWe used free or pedicled TAP flaps in 7 patients from Aug 2006 to April 2007, The age ranged from 7 to 42 years old, the perforator arteries was detected and labeled with a hand held Doppler flowmeter, the size of flaps ranged from 6 cm x 9 cm - 12 cm x 16 cm, the flaps designed with perforator artery included, all the flaps are based on the first perforator artery.

RESULTSAll the flaps survived well, no complication occurred with lowest donor site morbidity.

CONCLUSIONSThe thoracodorsal artery flap with latissimus dorsal muscle saved is a thin and reliable flaps with robust of blood supply, the flap can reduce significantly donor site morbidity and is a good choice for reconstructive surgery.

Adolescent ; Adult ; Back ; blood supply ; Child ; Female ; Humans ; Male ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; blood supply ; Thoracic Arteries ; transplantation ; Young Adult