OBJECTIVESTo investigate whether antisense human telomerase reverse transcriptase (hTERT) could inhibit the activity of telomerase and the proliferation of K562 cells.

METHODSThe antisense plasmid was constructed by reverse insertion of hTERT PCR product into plasmid pLNCX-neo. Then the constructed plasmid was introduced into K562 cells by liposomes-mediated DNA transfection. The inhibition effects of telomerase on the proliferation of K562 cells were analyzed by MTT and colony formation assay, the telomerase activity of K562 cells by TRAP-PCR ELISA methods.

RESULTSThe growth rate of antisense hTERT transfected K562 cells was significantly lower than those of the controls, and the colony formation capacity of the transfected cells decreased significantly (P < 0.01), the colony number is (100.33 +/- 7.57)/10(3) cells, (92.67 +/- 5.86)/10(3) cells and (50.33 +/- 6.11)/10(3) cells for control K562 cells, K562 neo cells and antisense hTERT transfected HL60 cells, respectively. The telomerase activity of antisense hTERT transfected K562 cells was significantly inhibited.

CONCLUSIONThe expression of an antisense sequence to the mRNA sequence of telomerase protein subunit can inhibit the activity of telomerase, slow the cell growth and inhibit the capacity of colony formation of K562 cells.

Cell Division ; drug effects ; Humans ; K562 Cells ; Plasmids ; genetics ; RNA, Antisense ; genetics ; pharmacology ; RNA, Messenger ; genetics ; Telomerase ; drug effects ; genetics ; metabolism ; Transfection

Aim To obtain the active components and targets of ginseng in the prevention and treatment of traumatic stress disorder(PTSD) through the method of network pharmacology. Methods The active components and target information of ginseng with medicinal value were obtained by TCMSP research platform, and the gene information closely related to the pathogenesis of PTSD was obtained by searching GeneCard and OMIM database. The two were matched to obtain the medicinal components and target genes of ginseng in the prevention and treatment of PTSD. The drug-dis- ease-target network diagram was drawn by R and Perl computer languages, and the target genes were analyzed by PPI network analysis, gene ontology ( GO ) and signal transduction pathway ( KEGG) enrichment analysis. Results According to the general pharmacological research methods of traditional Chinese medi cine, the screening parameters of active components were set, and nine kinds of high value medicinal ingredients of Panax ginseng were obtained. There was a drug-target relationship between the nine medicinal components and sixteen target genes related to PTSD disease. Through PPI, GO and KEGG analysis, it was found that the target genes were mainly enriched in physiological functions such as neurotransmitters, syn-aptic plasticity, ion channels and so on. Conclusions Ginseng has the pharmacological effect of preventing and treating PTSD, which may play a role in regulating the metabolism and receptor activity of monoamine neurotransmitters.

OBJECTIVETo investigate the inhibition effect of leukemic bone marrow stromal cells (BMSCs) on daunorubicin (DNR) induced apoptosis of human Jurkat cell line, and analyze the differentially expressed genes between Jurkat cells cocultured with leukemic BMSCs or without.

METHODSSuppression subtractive hybridization (SSH) was employed to establish subtracted cDNA library of differentially expressed genes in Jurkat cells cocultured with leukemic BMSCs and DNR. The cDNA fragments were sequenced and analyzed.

RESULTSThe differentially expressed gene cDNA library was successfully developed. Primary screening was done by reverse Northern hybridization. Thirty up-regulated and 22 down-regulated cDNA fragments were isolated and sequenced. Analysis and comparison were performed in GenBank using BLAST. These genes are related to cell cycle regulation, cell apoptosis and energy metabolism.

CONCLUSIONLeukemic BMSCs influence gene expression of Jurkat cells. The resulting differentially expressed genes might be associated with the protection of leukemic cells by BMSCs from injury.

Apoptosis ; drug effects ; Bone Marrow Cells ; drug effects ; metabolism ; pathology ; Cells, Cultured ; Coculture Techniques ; Daunorubicin ; pharmacology ; Gene Expression Profiling ; Gene Expression Regulation, Leukemic ; drug effects ; Gene Library ; Humans ; Jurkat Cells ; Stromal Cells ; drug effects ; metabolism ; pathology

BACKGROUNDBmi-1 gene determines the proliferative capacity of normal and leukemia stem cells. Expression of Bmi-1 has been found in all types of myeloid leukemia cells in both humans and mice. This study aimed at assessing the effect of antisense Bmi-1 expression on K562 cells proliferation and p16 protein (p16) expression.

METHODSA transcriptional repressor, Bmi-1 cDNA was cloned by reverse transcriptase polymerase chain reaction (RT-PCR) of its mRNA from K562 cells. A plasmid expressing antisense Bmi-1 mRNA was then constructed by reverse design of PCR primers and cloned to the plasmid pLNCX2; G418 was added to the medium after the plasmid was successfully introduced in K562 cells by lipofectin-mediated DNA transfection. The effects of the antisense expression on the proliferation of K562 cells were analyzed by using microculture tetrazolium and colony forming. Cell cycle was analyzed by using flow cytometry. The p16 expression of K562 cells was observed by immunofluorescence histochemical stain.

RESULTSK562 cells transfected with antisense Bmi-1 plasmid grew significantly slower than that of controls (the parental K562 and cells transfected with empty plasmid). The colony forming ability of antisense Bmi-1 plasmid transfected cells decreased significantly (P < 0.01) compared with controls. The p16 expression of cells transfected with antisense Bmi-1 was upgraded more apparently than that of controls.

CONCLUSIONThe antisense Bmi-1 gene can inhibit the growth of K562 cell and upgrade expression of p16 in K562 cells.

Cell Cycle ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p16 ; analysis ; Humans ; K562 Cells ; Nuclear Proteins ; antagonists & inhibitors ; genetics ; Plasmids ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; antagonists & inhibitors ; genetics ; RNA, Antisense ; physiology ; Repressor Proteins ; antagonists & inhibitors ; genetics

OBJECTIVETo explore the reliability and validity of digital pulse wave analyzing method in evaluating arterial compliance in population-based study.

METHODSAll 415 adults (132 men and 283 women) aged 20 to 86 years old were selected from urban (220 persons) and rural (195 persons) areas, respectively by a stratified randomly sampling method. Arterial compliance, evaluated by stiffness index (SI), was measured by using digital pulse wave analyzing method from the Pulse trace machine (Micro medical, London), and the SI value was determined accordingly.

RESULTSIn the study on both repeatability and stability, there was a perfect correlation between the frequent measurements for one individual either on one occasion or on two 40-days-apart occasions. The SI values were not significantly different between the urban and the rural, men and women. Multiple stepwise regressions showed that systolic blood pressure and age were positively correlated with SI value, respectively (both P values were less than 0.001). The correlation kept unchanged after taking account of gender, BMI and heart rate. The mean SI values for people aged 20 to 29, 30 to 39, 40 to 49, 50 to 59 and > or = 60 were 7.35, 8.84, 10.41, 10.95 and 12.01 m/s (P < 0.01), respectively.

CONCLUSIONSBoth systolic blood pressure and age should be contributed as the main influencing factors of arterial compliance. Digital pulse wave analyzing method is a preferable measurement in evaluating arterial compliance in population-based study due to its better repeatability and stability.

Adult ; Aged ; Aged, 80 and over ; Arteries ; physiopathology ; Arteriosclerosis ; epidemiology ; physiopathology ; China ; epidemiology ; Compliance ; Female ; Humans ; Male ; Middle Aged ; Pulse ; methods ; Rural Population ; Sampling Studies ; Urban Population

OBJECTIVETo investigate the surgical treatment for hilar cholangiocarcinoma(HCCA) of Bismuth-Corlette type IV.

METHODSThe clinical data of 73 HCCA patients admitted in Southwest Hospital, the Third Military Medical University from January 2002 to December 2008 were analyzed retrospectively. There were 41 males and 32 females, aged from 30 to 84 years old (averaged, 56.8 years old). All patients were diagnosed as hilar mass with hilar biliary obstruction by B-ultrasound, CT, magnetic resonance imaging, endoscopic retrograde cholangiopancreatography, magnetic resonance cholangiopancreatography or percutaneous transhepatic cholangiography, and confirmed by pathological examination in intra-operation and post-operation. Diagnosis of all patients were according to Bismuth-Corlette type IV.

RESULTSFifteen cases underwent percutaneous transhepatic cholangial drainage, stents were implanted in 8 patients. Simple internal drainage was performed on 25 of the 73 patients and 4 with palliative resection. Radical resection was performed on 19 patients. The radical resection rate of HCCA were 26.0%. The 1, 3 years of survival rates were 36.8%, 10.5% respectively. The 1 year survival rate was 6.3% in drainage group.

CONCLUSIONRadical resection is the potentially curative treatment for HCCA, a sufficient, reasonable use of internal and external drainage would improve the patient's quality of life.

Adult ; Aged ; Aged, 80 and over ; Bile Duct Neoplasms ; diagnosis ; surgery ; Bile Ducts, Intrahepatic ; Cholangiocarcinoma ; diagnosis ; surgery ; Drainage ; Female ; Follow-Up Studies ; Hepatectomy ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Treatment Outcome

OBJECTIVEThe aims of this study were to evaluate whether the presence of -2518A/G polymorphism in the distal regulatory region of the monocyte chemotactic protein-1 (MCP-1) was associated with tuberculosis (TB) in Chongqing Han population and to find whether it has a significant impact on the pediatric patient.

METHODOne hundred children [ < or = 15 years old, mean age (7.3+/-4.6) years, 53 male, 47 female] and one hundred adults [51 male, 49 female, age (44.6+/-13.5) years with TB] and 200 healthy controls of comparable age were screened for genotype by PCR-sequence-specific primer (SSP) method. MCP-1 levels in the sera were detected by ELISA.

RESULT(1) TB patients and controls showed different single nucleotide polymorphism (SNP) distribution patterns (58%, 36%). MCP-1 alleles -2518G was associated with increased TB susceptibility (P<0.01). (2) The -2518 GG genotypes was associated with increased TB susceptibility (32% in TB patients and 13% in non-TB controls respectively, P<0.01). (3) The odds of developing TB in genotypes GG were higher than those in homozygous AA, and the risk was higher in children than in adult (7.0-fold in children and 5.1-fold in adults, respectively). (4) Cases of homozygous GG had the highest plasma levels of MCP-1, which increased the likelihood of developing TB. Furthermore, higher levels were observed in children than in adults.

CONCLUSIONThese findings suggest that persons bearing the MCP-1 genotype GG produce high concentrations of MCP-1, which increases the risk of active TB infection in Chongqing Han people. These findings are more significant in child patients than in adult patients with TB.

Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Chemokine CCL2 ; blood ; genetics ; Child ; Child, Preschool ; DNA Primers ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Tuberculosis ; ethnology ; genetics ; metabolism

Objective To study a method for monitoring multichannel electrode-scalp contact impedances in electrical impedance tomography (EIT), and to provide support for exploring the impact of contact impedances on imaging and the suppression method.Methods A reference electrode was introduced for the system,placed on the back of the neck or the top of the head. In the spare time of boundary voltage signal acquisition phase, the method of two electrodes was adopted to measure the electrical impedance between each imaging electrode and reference electrode, which was used to analyze the characteristics of multichannel contact impedances.Based on the existing EIT hardware system,the module for monitoring multichannel contact impedance was implemented.Results The calibration board test showed that the measurement error of this method was less than 1‰,and the maximum relative fluctuation of the measurement result within 10 hours was less than 1‰. Preliminary human test indicated that this method had the ability to obtain two kinds of information about spatial consistency and temporal stability of 16-channel contact impedances. Conclusion This method has high accuracy and stability,so it meets the requirements of monitoring multichannel contact impedances in brain EIT.

Objective:To discuss the efficacy of Bushen Huoxue decoction for recovery in patients with intra uterine adhesions （IUA） after trans cervical resection adhesions （TCRA） operation， and to investigate its effect on fibrosis factor. Method:The 110 patients were randomly divided into two groups by random number table （55 cases for control group and 55 cases for observation group）. The patients in both groups got TCRA， and after the operation， they got estradiol valerate tablets+progesterone capsules. The patients in control group additionally got Jinfengwan before meals， 10 g/time， 2 times/day. Patients in observation additionally got syndrome differentiation and treatment by Bushen Huoxuetang， 1 dose/day. The treatment course was 3 months in both groups， and 6 months' follow-up was recorded. Before and after treatment， menstrual volume， period and cycle were recorded. During the follow-up， pregnancy， ectopic pregnancy and abortion were also recorded. Endoscopy and vagina color Doppler ultrasound were conducted to detect endometrial thickness， uterine volume， resistance index （RI）， pulsation index （PI） and blood flow index （FI）. Scores of IUA and syndrome of kidney deficiency and blood stasis were graded. At the 6^th months after treatment， rate of intrauterine re-adhesion was recorded. Levels of transforming growth factor-β₁ （TGF-β₁）， connective tissue growth factor （CTGF）， matrix metalloproteinase inhibitor-1 （TIMP-1）， platelet-derived growth factor （PDGF）， insulin-like growth factor-1 （IGF-1） and matrix metalloproteinase-9 （MMP-9） were detected， and safety was evaluated. Result:Total effective rate of clinical efficacy was 96.36% （53/55） in observation group， higher than 83.64% （46/55） in control group （χ²=4.959， P<0.05）. Recovery rate of menstrual volume， period and cycle were 92.73% （51/55）， 90.91% （50/55）， and 94.55% （52/55） in observation group， higher than 72.73% （40/55）， 76.36% （42/55）， and 76.36% （42/55） in control group（χ²=7.698，χ²=4.245，χ²=7.313，P<0.05， P<0.01）. During the follow-up， rate of intrauterine re-adhesions was 23.64% （13/55） in observation group， lower than 43.64% （24/55） in control group （χ²=4.927，P<0.05）. Pregnancy rate was 45.45% （25/55）， higher than 25.45% （14/55） in control group（χ²=4.807，P<0.05）. Score of syndrome of kidney deficiency and blood stasis and IUA scores in observation group were lower than those in control group （P<0.01）. Endometrial thickness， uterine volume， and FI were higher than those in control group （P<0.01）. Levels of RI， PI， TGF-β₁， CTGF， PDGF， IGF-1 and TIMP-1 were lower than those in control group （P<0.01）， while level of MMP-9 was higher than that in control group （P<0.01）. No adverse reactions related to traditional Chinese medicine were found. Conclusion:Bushen Huoxue decoction can promote the recovery of menstruation， increase the thickness of endometrium， improve the blood circulation of uterus， regulate the expression of fibrogenic factors in vivo， prevent and treat intrauterine adhesions， reduce the rate of intrauterine re-adhesions， and improve the rate of pregnancy， with a good comprehensive effect and high safety for clinical use.

Objective To examine the effect of low temperature on trehalose and trehalase levels in Culex pipiens pallens. Methods The fourth instar larvae and female adult mosquitoes of Cx. pipiens pallens were exposed at 4 ℃ for 0, 1, 3, 6, 12, 18, 24 h and 0, 1, 3, 6, 12, 18, 24, 36, 48, 72 h, respectively. Then, the trehalose and trehalase contents were detected by enzyme-linked immunosorbent assay (ELISA) in mosquitoes. Results The contents of trehalose and trehalase significantly increased in the larval and female adult mosquitoes post-exposure to low temperature. The changing trend of trehalose levels was consistent in the larval and female adult mosquitoes, and the highest levels were (2.458 8 ± 0.379 2) mg/g and (2.825 7 ± 0.211 1) mg/g 3 h post-exposure to low temperature, respectively. The trehalose and trehalase levels fluctuated greatly within the first 6 h post-exposure to low temperature. Following adaptation for a period of time, the trehalose and trehalase levels remained at a relatively high level. Conclusion Low temperature may induce the production of trehalose and trehalase in Cx. pipiens pallens, and the trehalose and trehalase may play an important role in the improvement of the cold resistance.