7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide ; toxicity ; Carcinogens ; toxicity ; Cell Line, Tumor ; DNA Damage ; drug effects ; Dose-Response Relationship, Drug ; HSP70 Heat-Shock Proteins ; biosynthesis ; Humans

Bletilla striata has been used as traditional Chinese medicine for several centuries. In recent years, the quality and quantity of wild B. striata plants have declined sharply due to habitat deterioration and human over-exploitation. Therefore, it is of great urgency to evaluate and protect B. striata wild plant resource. In this study, sequence-related amplified polymorphism (SRAP) markers were applied to assess the level and pattern of genetic diversity in twelve populations of B. striata. The results showed a high level of genetic diversity (PPB = 90.48%, H = 0.349 4, I = 0.509 6) and moderate genetic differentiation among populations (G(st) = 0.260 9). Based on the unweighted pair-group method with arithmetic average (UPGMA), twelve populations gathered in three clusters. The cluster 1 included four populations. There are Nanjing, Zhenjiang, Xuancheng and Hangzhou. The seven populations which come from Hubei Province, Hunan Province, Jiangxi Province and Guizhou Province belonged to the cluster 2. The cluster 3 only contained Wenshan population. Moreover, Mantel test revealed significant positive correlation between genetic distances and geographic distances (r = 0.632 9; P < 0.000 1). According to the results, we proposed a series of conservation consideration for B. striata.
China ; Genetic Markers ; Genetic Variation ; Genetics, Population ; Orchidaceae ; genetics ; Phylogeny ; Plants, Medicinal ; genetics

Objective To study the effects of a new phosphodiesterase inhibitor MS23 on the myogenic responses of diabetic rat coronary artery (RCA) and the myocyte potassium currents via the voltage-gated potassium channels (Kv).Methods Rats were rendered diabetic with a peritoneal injection of streptozocin at 60 mg · kg-1.At the 4th week after treatment,the diabetic rats were randomly divided into model group,low-dose,and high-dose groups of MS23 (0.3 mg · kg-1 and 1.0 mg · kg-1 orally administrated once a day for 8 weeks),while the normal rats were set as the control group.Model group and control group were fed with 0.9％ NaCl of equal volume once a day from the fourth week to the twelfth week.A wire myograph was used to observe the myogenic effects of isolated RCA.The Kv currents of single rat coronary arterial smooth muscle cell (RCASMC) were studied with patch clamp.Results After administration,the maximum vasorelaxations of RCA to acetylcholine in control group,model group,low-dose,high-dose group were (74.45 ± 6.30) ％,(56.84 ± 5.51) ％,(77.52 ± 5.27) ％,(86.33 ± 8.88) ％,respectively.The maximum vasorelaxations to sodium nitroprusside in the four groups were (88.50 ± 5.81)％,(56.65 ±8.62)％,(80.50 ±7.85)％,(91.60 ±5.97)％,respectively.The maximum RCASMC Kv current density in the four groups were (53.69±4.02) pA · pF-1,(28.25±3.69) pA · pF-1,(38.78±4.55) pA · pF-1,(63.35 ± 4.57) pA · pF-1,respectively.There were significant differences between the model group and the blank group/low-dose group/high-dose group (P ＜ 0.05).The maximum vasorelaxations of diabetic RCA to MS23 (30 μmol · L-1) was (70.69 ± 12.72)％,and preincubation with Kv inhibitor 4-AP (3 mmol · L-1) reduced the MS23-induced maximum relaxation to (34.47 ± 10.08) ％,with significant difference (P ＜ 0.05).Conclusion MS23 relaxes diabetic rat coronary artery and enhances diabetic RCASMC Kv currents.

OBJECTIVETo observe the effect of nitrotyrosine on renal expressions of nuclear factor-κB (NF-κB), monocyte chemoattractant protein-1 (MCP-1), and transforming growth factor-β1 (TGF-β1) in rats with diabetic nephropathy (DN).

METHODSRat DN models established by a single intraperitoneal injection of streptozotocin (STZ) were randomly allocated into model group, nitrotyrosine group and ebselen group, with untreated rats as the normal control group. The rats were given the corresponding drugs for 8 weeks, and after the last administration, the 24-h urinary protein level was measured and the kidneys of the rats were harvested for detecting the protein and mRNA expressions of NF-κB, MCP-1 and TGF-β1 with immunohistochemistry and RT-PCR, respectively. The pathological changes of the kidneys were assessed microscopically.

RESULTSCompared with those in the model group, the 24-h urinary protein level and expressions of NF-κB, MCP-1 and TGF-β1 mRNA and protein in the renal tissues were significantly increased by nitrotyrosine treatment, which also caused worsened renal pathology, while treatment with ebselen significantly ameliorated these changes.

CONCLUSIONNitrotyrosine can up-regulate the mRNA and protein expressions of NF-κB, MCP-1 and TGF-β1 and aggravate the inflammatory reaction in the renal tissue of DN rats to promote the progression of DN.

Animals ; Chemokine CCL2 ; metabolism ; Diabetes Mellitus, Experimental ; Diabetic Nephropathies ; metabolism ; Male ; NF-kappa B ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism ; Tyrosine ; analogs & derivatives ; pharmacology

Arginine vasopressin (AVP), a neurohormone and hemodynamic factor implicated in the pathophysiology of hypertension and congestive heart failure, can also act as a growth-stimulating factor. Our previous work demonstrated that AVP is a mitogen for neonatal rat cardiac fibroblasts (CFs). In the present study, we extended our investigations to adult rat CFs to explore whether AVP could induce adult rat CF proliferation and, if so, to identify the mechanism involved. Adult rat CFs were isolated, cultured and subjected to AVP treatment. DNA synthesis and cell cycle distribution were analyzed by [(3)H]-thymidine incorporation and flow cytometry. Cellular extracellular signal-regulated kinase 1/2 (ERK1/2) activity was measured by in vitro kinase assay using myelin basic protein (MBP) as a substrate. Protein expressions of total- and phospho-ERK1/2, p27(Kip1), cyclins D1, A, E were assessed by Western blot. The results showed that AVP stimulated DNA synthesis in adult rat CFs, and the effect was abolished by a V1 receptor antagonist, d(CH(2))(5)[Tyr(2)(Me), Arg(8)]-vasopressin (0.1 μmol/L), but not by a V2 receptor antagonist, desglycinamide-[d(CH(2))(5), D-Ile(2), Ile(4), Arg8]-vasopressin (0.1 μmol/L). AVP induced an activation of ERK1/2, which could be mimicked by the protein kinase C (PKC) activator, phorbol 12-myristate 13-acetate (PMA, 30 nmol/L, 5 min), but abolished by depletion of PKC via chronic PMA incubation (2.5 μmol/L, 24 h). In addition, AVP down-regulated protein expression of p27(Kip1), increased protein expressions of cyclins D1, A and E, and induced cell cycle progression from G(0)/G(1) into S stage. Inhibition of ERK1/2 activation by PD98059 (30 μmol/L) abolished the effect of AVP on DNA synthesis, protein expressions of p27(Kip1), cyclins D1, A and E as well as cell cycle progression. These results suggest that AVP is also a growth factor for adult rat CFs. The mitogenic effect of AVP is mediated via V1 receptors and PKC-ERK1/2 pathway. Moreover, AVP modulates the expressions of cell cycle regulatory proteins p27(Kip1) and cyclins D1, A and E, which lie downstream of ERK1/2 activation, and induces cell cycle progression in adult rat CFs.
Animals ; Antidiuretic Hormone Receptor Antagonists ; pharmacology ; Arginine Vasopressin ; pharmacology ; Cell Cycle ; Cell Cycle Proteins ; metabolism ; Cell Proliferation ; Fibroblasts ; cytology ; drug effects ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Myocardium ; cytology ; Phosphorylation ; Protein Kinase C ; metabolism ; Rats ; Signal Transduction ; Tetradecanoylphorbol Acetate ; pharmacology

OBJECTIVEAIM To establish a drug screening model based on transcriptional regulation of estrogen responsive element (ERE) and use it to screen compounds for discovering new ligands of estrogen receptor (ER) subtypes.

METHODA recombinant reporter vector pERE-TAL-SEAP was constructed by inserting a synthetic sequence composed of five tandem copies of EREs upstream of promoter of the reporter vector pTAL-SEAP. The pERE-TAL-SEAP and the internal control plasmid pCMV were transiently co-transfected into Hela cells expressing ER subtype or ER subtype, and the effects of pure ER agonists 17estradiol, phytoestrogen genistein and pure ER antagonist ICI182, 780 on reporter gene SEAP expression were observed.

RESULTIn the Hela cells expressing ER alpha or ER beta subtype, the expression of SEAP gene were induced in a dose dependent manner by 17-estrodiol with a maximal effect at approximately 10 nmol.L-1 and with EC50 of (80.58 +/- 8.51) pmol.L-1 and (103.90 +/- 5.29) pmol.L-1, respectively, so done by phytoestrogen genistein with a maximal effect at 1 mumol.L-1 and with EC50 of (10.86 +/- 0.75) nmol.L-1 and (39.38 +/- 2.26) nmol.L-1, respectively. The maximal level induced by estrodiol and genistein were about 7-14 fold higher than that of vehicle. The pure antiestrogen ICI182, 780 at concentration of 1 mumol.L-1 completely blocked the inductions of 17-estrodiol and genistein.

CONCLUSIONThe cellular drug screening model can be established by transfecting reporter vector pERE-TAL-SEAP in Hela cell lines expressing ER alpha or ER beta. The cell lines can be used to screen compounds with estrogenicity by testing SEAP activity in the culture media of cells growing in microtitier wells. The system should provide an efficient model for screening and analyzing the activity of large numbers of ligands of ER.

Drug Evaluation, Preclinical ; methods ; Estradiol ; pharmacology ; Estrogen Receptor alpha ; Estrogen Receptor beta ; Gene Expression Regulation ; drug effects ; Genes, Reporter ; Genistein ; pharmacology ; HeLa Cells ; Humans ; Ligands ; Promoter Regions, Genetic ; Receptors, Estrogen ; genetics ; Transfection

OBJECTIVETo investigate the effect of acupuncture on blood oxygen saturation in the patient of obstructive sleep apnea-hypopnea syndrome (OSAHS) in sleeping and to evaluate the therapeutic effect of acupuncture on this disease.

METHODSThirty cases with OSAHS were treated with acupuncture at Shanglianquan (Extra), Fengfu (GV 16), Yamen (GV 15), Fengchi (GB 20), etc. 3-5 sessions each week. After treatment of 30 sessions, apneahypopnea index (AHI), mean blood oxygen saturation (MSaO2), the lowest blood oxygen saturation (LSaO2), oxygen desaturation > or = 4% index (ODI4), the mean blood oxygen saturation of oxygen desaturation when SaO2 < 90%, the longest time of oxygen saturation > or = 4% were observed before and after treatment.

RESULTSThe effective rate of acupuncture was 23.3% for OSAHS. After acupuncture, AHI and ODI4 significantly reduced (P < 0.01); LSaO2 significantly increased (P < 0.01); MSaO2 and the mean blood oxygen saturation of oxygen desaturation when SaO2 < 90% significantly enhanced (P < 0.05); the longest time of oxygen saturation > or = 4% did not significantly change.

CONCLUSIONThe acupuncture treatment has intervenient effect on OSAHS and alleviates anoxia, so acupuncture is one of therapies improving anoxia in patients of OSAHS.

Acupuncture Therapy ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Oxygen ; blood ; Sleep Apnea, Obstructive ; blood ; therapy

OBJECTIVETo investigate the epidemiology, peritoneal dialysis (PD) related complications and survival outcomes of 236 patients with end-stage renal disease (ESRD) undergoing continuous ambulatory peritoneal dialysis (CAPD) in our center from January, 2004 to November, 2009.

METHODSThe data including patient gender, age, time of PD initiation, addresses, types of medical reimbursement, primary diseases, modes of PD catheter placement surgery, types of PD catheter, PD-related complications, and time of drop out were retrospectively analyzed. PD catheter migration rate, peritonitis rate, drop out rate (DOR), length of the time of PD therapy (TOT), and survival rate were calculated and compared with those of patients in other PD centers.

RESULTSThe number of newly introduced patients increased gradually in the years from 2004 to 2009. The mean age of newly introduced patients was 47-/+16 years, and patients with age below 60 years accounted for 77.96%. Patients who paid for their own expenses accounted for 67.37% of all, and the rate of these patients decreased gradually. Similar to that in Asian-Pacific region, chronic glomerulonephritis was the most frequent cause of ESRD followed by diabetic nephropathy. The number of patients with chronic glomerulonephritis or obstructive nephropathy as the primary diseases was greater in this center than that reported in the Asian-Pacific region, accounting for 54.66% and 11.02% of all patients, respectively. In contrast, the patients with diabetic nephropathy or benign arteriolar renal sclerosis were less, accounting for 12.29% and 10.17% of all, respectively. PD catheter migration rate (8.05%) and peritonitis rate (1:44.22 patient-months) were both lower than those reported. The patient survival rates at 1, 2, 3 years were 83.65%, 51.59% and 29.81%, respectively, lower than those of other centers in the developed countries but higher than the mean levels in China. DOR decreased gradually to 11.56% in 2009, and TOT increased to 23.61 months.

CONCLUSIONThe above characteristics of the patients are related to many factors, including the "PD first" principle, high prevalence of urinary calculosis in the primary source regions of most patients, preventive partial omentum resection in some patients, education and follow-up for patients, and increased expense cover by medical insurance.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Glomerulonephritis ; complications ; Humans ; Kidney Failure, Chronic ; etiology ; therapy ; Male ; Middle Aged ; Peritoneal Dialysis, Continuous Ambulatory ; adverse effects ; statistics & numerical data ; Retrospective Studies ; Survival Rate ; Young Adult

BACKGROUNDPrimary biliary cirrhosis (PBC) is a chronic and slowly progressive cholestatic liver disease characterized by destruction of the interlobular bile ducts and a striking female predominance. The aim of this study was to identify associations between estrogen receptor (ESR) gene polymorphisms with the risk of developing PBC and abnormal serum liver tests in a Chinese population.

METHODSThirty-six patients with PBC (case group) and 35 healthy individuals (control group) from the First Hospital of Jilin University were studied. Whole genomic DNA was extracted from all the participants. Three single-nucleotide polymorphisms (rs2234693, rs2228480, and rs3798577) from ESR1 and two (rs1256030 and rs1048315) from ESR2 were analyzed by a pyrosequencing method. Demographic data and liver biochemical data were collected.

RESULTSSubjects with the T allele at ESR2 rs1256030 had 1.5 times higher risk of developing PBC than those with the C allele (odds ratio [OR] = 2.1277, 95% confidence interval [CI] = 1.1872-4.5517). Haplotypes TGC of ESR1 rs2234693, rs2228480, and rs3798577 were risk factors for having PBC. The C allele at ESR1 rs2234693 was associated with abnormal alkaline phosphatase (OR = 5.2469, 95% CI = 1.3704-20.0895) and gamma-glutamyl transferase (OR = 3.4286, 95% CI = 1.0083-13.6578) levels in PBC patients.

CONCLUSIONSESR2 rs1256030 T allele may be a significant risk factor for the development of PBC. Screening for patients with gene polymorphisms may help to make early diagnoses in patients with PBC.

Adult ; Aged ; Asian Continental Ancestry Group ; Case-Control Studies ; Estrogen Receptor alpha ; genetics ; Estrogen Receptor beta ; genetics ; Female ; Gene Frequency ; genetics ; Genetic Predisposition to Disease ; genetics ; Haplotypes ; genetics ; Humans ; Liver Cirrhosis, Biliary ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; genetics ; Receptors, Estrogen ; genetics

Objective To compare the function recovery of multiple injuries combined with floating knee joint injury and simple knee joint injury,and to analyze the risk factors.Methods A retrospective case control study was conducted to analyze the clinical data of 41 patients with multiple injuries combined with Blake and McBryde Ⅱ A floating knee injury admitted to Xijing Hospital of Air Force Medical University from June 2011 to June 2017.There were 26 males and 15 females,aged 18-76 years,with an average of 34.5 years.There were 25 patients with simple knee joint injury and and 16 patients with multi-joint combined injury involving knee joint and ipsilateral hip joint or ankle joint injury.Surgical fixation was performed in different parts by external fixation,intramedullary nail and plate screw fixation.According to the Kalstr(o)m and Olerud functional evaluation criteria,the excellent and good rate of postoperative functional recovery was compared between the two groups.The surgical fixation methods of the two groups were compared.Logistic regression analysis was performed on the influencing factors of functional recovery.Results The patients were followed up for 1-7 years,with an average of 3 years.The excellent and good rate of overall functional recovery in the two groups was 68％,and the rate was 84％ in simple knee injury group and 44％ in multi-joint combined injury group (P ＜ 0.01).There was no significant difference in the ratio of intramedullary nail and plate screw fixation between the two groups (P ＞ 0.05),while the proportion of the external fixation in the multi-joint combined injury group [31％ (5/16)] was significantly higher than that in the simple knee joint injury group [16％ (4/25)] (P ＜ 0.05).Logistic regression analysis showed that the rate of external fixation was an independent factor affecting the postoperative function (OR =0.15,P ＜ 0.01).Conclusions The postoperative function in multi-joint injury patients is poorer than in the single joint injury patients.The higher rate of using external fixation in multi-joint injury patients is a risk factor.For Blake and McBryde Ⅱ A floating knee injury combined with multi-joint injury,less external fixation should be used,so as to improve the postoperative function.