Background and purpose: Cyclin-dependent kinase 4 (CDK4) is a kind of protein kinases regulating the cell cycle progression, which has been reported to be overexpressed in endometrial carcinoma tissues. But the role of CDK4 in endometrial carcinogenesis and relative mechanisms has not been identiifed yet. In this study, we used a small interfering RNA targeting CDK4, and explored the effects of CDK4 on endometrial cancer cells HEC-1B biological function and relative mechanisms.Methods:The chemically synthesized small interfering RNA targeting CDK4 (si-CDK4) was transiently transfected into HEC-1B cells;the quantitative real time-PCR assays and Western blot assays were performed to explore the mRNA and protein expression levels of CDK4 and its downstream genes, Rb and p-Rb, in HEC-1B cells upon transfection;Moreover, the CCK-8, lfow cytometry (FCM) and invasion assays were performed to indentify the effects of si-CDK4 on the proliferation, cell cycle distribution, apoptosis and invasion abilities of HEC-1B cells, respectively. Results:The results showed that the mRNA and protein expressions of CDK4 were suppressed in HEC-1B cells upon transfection with si-CDK4 (P<0.01);Suppression of CDK4 inhibited cell proliferation and invasion of HEC-1B cells;the number of cells migrating through the transwell membrane in si-CDK4 group was 117±21, which was much fewer than the cells in si-control (269±39) and untreated groups (262±35) (P<0.01);the early apoptosis rate of cells treated with si-CDK4 [(21.7±3.5)%] was much higher than the untreated [(12.4±2.1)%] and si-control groups [(11.8±1.9)%] (P<0.01);moreover, suppression of CDK4 increased cells in G1 phase (P<0.01) and correspondingly decreased cells in S phase (P<0.01);further Western blot results showed that suppression of CDK4 down-regulated the expression of p-Rb in cells, but did not inlfuence the expression of total Rb. Conclusion:CDK4-siRNA speciifcally and efifciently blocks the constitutively activated CDK4 in human endometrial cancer cells HEC-1B, resulting in tumor suppression.

Tissues and organs generate angiogenesis under the stimulation of angiogenic factors in physiological or pathological conditions.Multiple signal pathways including VEGF, Notch, Wnt/β-catenin, Ang1(2)/tie2 and PIK-Akt etc.have effects on various stages of angiogenesis.VEGF exerts irreplaceable effects on the whole process of angiogenesis through multiple signal pathways.Over the past few years, new progress has been made in the researches of mechanisms regulating angiogenesis through VEGF-related signal pathways both at home and abroad.These findings provide us new theoretical basis for clarification of the pathogenesis of many diseases and clinical drug development.In this article we will summarize the recent research progress in this field, hoping to provide new possibilities for the treatment of angiogenesis-related diseases.

Objective To analyze factors affecting quality of life (QOL)in patients with chronic hepatitis B (CHB),and provide reference for improving QOL of patients. Methods The MOS 36-item short form health sur-vey (SF-36)was adopted to survey QOL of patients with CHB,Morisky medication adherence scale was used to measure patients'adherence to medication,factors affecting QOL of patients with CHB were analyzed. Results Of 357 CHB patients,271(75.91% )were married,107(29.97% )received college or above education,163(45.66% ) patients'average household monthly income were ￥ 2000-￥ 5000,138(38.66% )patients'family members were also with CHB,198 (55.46% )patients smoked,150 (42.02% )drank. The average score of CHB patients' adherence to medication was (2.15±1.29). Factors affecting QOL in patients with CHB were age,education level, duration of disease,whether or not hospitalized,whether or not drink,as well as adherence to medication. Age, drink,and duration of hepatitis B,and previous hospitalization were negative factors affecting QOL in patients with CHB,education level and adherence to medication were positive factors affecting QOL in patients with CHB. Conclusion Strengthening CHB patients'understanding on disease and improving their medication adherence can help them to improve QOL.

Proliferation and differentiation of neural stem cells is regulated by autologous or external, adjacent or remote cell signaling pathways. This paper reviewed the studies about the Notch, BMP, Wnt, Shh signaling pathways related to brain neurogenesis.

Objective To establish a quick method to analyze vinorelbine ( NVB) in plasma of beagle dogs and study its pharmacokinetics of long-circulation and thermosensitive liposome loaded vinorelbine bitartrate.Methods The plasma was treated with liquid-liquid extraction after precipitation in methanol.The analysis was perfomed on a Venusil XBP C18 column(2.1 mm ×50 mm, 3 μm) at 35℃,the mobile phase consisted of methanol and water( containing 10 mmol/L ammonium acetate, 1%acetonitrile) 80∶20 and injection volume was 10μl.The type of mass spectrum was multireactive monitoring(MRM) in a positive mode.The monitor transitions were m/z 779.4-765.4 for vinorelbine and m/z 825.4-122 for vincristine.Results The concentration range from 10 to 2500 ng/ml had a good linearity ( r=0.0994).The precision, accuracy and extraction efficiency were acceptable.The plasma samples were stable for 10 days at -20℃ and 24 h at room temperature.Pharmacokinetic study in beagle dogs showed that the main parameters for injection and liposome were Cmax(833.51 ±150.42) and (1397.95 ±443.05)ng/ml, AUC(0-t) (577.16 ±223.57) and (1059.82 ±408.27) ng/ml· h, Cl(3014.64 ±1049.17)and 1633.10 ±551.77 ml/(h· kg), respectively.Conclusion A reliable HPLC-MS/MS method for vinorelbine analysis is established and can be applied to the pharmacokinetics study of liposome.The results show that liposome has a higher AUC, Cmax and longer Cl than injection.Meanwhile, liposome has a lower irritability.

Objective To explore the expressions and clinical significances of deleted in liver cancer-1 (DLC-1 ) and Rho associated coiledcoil forming protein kinase (ROCK )Ⅰ in non-small lung cancer (NSCLC).Methods The expressions of DLC-1 and ROCKⅠ in NSCLC and adjacent tissue of 48 patients with pathologically confirmed as NSCLC and undergone surgical resection were detected by immunohistochemis-try EnVision method.The correlations among DLC-1 protein,ROCKⅠ protein and the clinical pathological characteristics were analyzed.The prognostic value of DLC-1 in patients with NSCLC was studied.Results The expression of DLC-1 protein in NSCLC tissue was low or missing,and the positive rate was 33.3%(16/48),significantly lower than that in the tissue adjacent to carcinoma 70.8% (34/48),with statistical significance (χ2 =13.523,P<0.01).The positive expression rate of ROCKⅠprotein in NSCLC was 58.3%(28/48),higher than that of tissue adjacent to carcinoma 0(0/48),with statistical significance (χ2 =39.529, P<0.01).The expression of DLC-1 protein was correlated with tumor differentiation,lymph node metastasis and clinical stage,rather than with sex,smoking history and organization type.Through the correlation analy-sis,the expression of ROCKⅠin DLC-1 positive group was 37.5%(6/16),and the expression rate of ROCKⅠ in DLC-1 negative group was 68.8%(22/32).There was negative correlation between DLC-1 and ROCKⅠin NSCLC tissues (r=-2.214,P=0.039).The 3 year survival rate in DLC-1 protein high expression group was obviously higher than that in low expression group,with statistical significance (P=0.043).Conclusion Low or missing expression of DLC-1 and high expression of ROCKⅠ protein may play an important role in the occurrence and development of NSCLC.Detecting the expression of DLC-1 and ROCKⅠprotein may be useful for evaluating the biological behavior and prognosis of NSCLC.

Objective To explore the effects of morroniside on the expression of Angiopoietin-1 (Ang-1) and Tie-2 in a rat after focal cerebral ischemia-reperfusion. Methods 20 male Sprague-Dawley rats were randomly divided into sham group (n=4), ischemia group (n=4), and morroniside groups (low, medium and high dosage groups, n=4). The middle cerebral artery were occluded for 30 min, and re-perfused. Morroniside was administered intragastrically once a day at dose of 30 mg/kg, 90 mg/kg and 270 mg/kg after operation. The expression of Ang-1 and Tie-2 in the ischemic ipsilateral cortex were detected with Western blotting analysis 7 days after operation. Results The expression of Ang-1 and Tie-2 increased in the ischemia group compared with the sham group (P<0.01), and both of them further increased in the morroniside groups of high dosage compared with the ischemia group (P<0.01), and the expression of Tie-2 also increased in the morroniside groups of medium dosage (P<0.001). Conclusion Morroniside could increase the expression of Ang-1 and Tie-2 in the ischemic ipsilateral cortex after ischemia-reperfusion in rats, suggesting promoting the angiogenesis after ischemia.

Objective To explore the effects of morroniside on the expression of CD34 in ipsilateral cortex of rats after focal cerebral isch-emia-reperfusion. Methods 45 male Sprague-Dawley rats were divided into sham group (n=9), ischemia group (n=9), and morroniside groups (low, medium and high dosage groups, n=9). The middle cerebral artery were occluded for 30 minutes, and reperfused. Morroniside was administered intragastrically once a day at dose of 30 mg/kg, 90 mg/kg, 270 mg/kg after operation. The expression of CD34 in the isch-emic ipsilateral cortex were detected with immunohistochemistry (n=6) and Western blotting (n=3) 7 days after operation. Results The ex-pression of CD34 increased in the ischemia group compared with the sham group, and further increased in the morroniside groups of high dos-age compared with the ischemia group (F>14.865, P<0.001). Conclusion Morroniside could increase the expression of CD34 in the ischemic ipsilateral cortex after ischemia-reperfusion in rats, which may promote the angiogenesis and neurogenesis after ischemia.

Objective To investigate the capsular polysaccharide (CPS) serotypes and molecular characteristics of carbapenem resistant hypermucoviscous Klebsiella pneumoniae (CR-HMKP) and study the possible mechanism of carbapenem resistance. Methods A retrospective study was conducted on 18 nonduplicate CR-HMKP strains which were collected from the First Affiliated Hospital of Nanchang University from 2012 to 2016. The clinical data were retrieved from medical records. The capsular serotypes, resistance genes and virulence factors were detected by polymerase chain reaction and DNA sequencing. Antimicrobial susceptibility testing was determined on VITEK 2 compact system. The CR-HMKP strains were characterized molecularly by using PCR, multilocus sequence typing (MLST) and pulsed field gel electrophoresis (PFGE). Modified carbapenem inactivation method was used to screen carbapenemase-producing strains. Plasmid conjugation transfer experiments were carried out to study transmission of carbapenem resistance. Results Eighteen (2.7％) CR-HMKP isolates were identified, which belonged to 4 serotypes, including wzi128-K1 (n=1), wzi206-K57 (n=1), wzi2-K2 (n=2), and wzi64-K14.64 (n=14). PCR and sequencing analysis identified blaNDM-1 gene in 2 CR-HMKP strains, blaKPC-2 gene in 17 strains, qnrS1 gene in 18 strains, blaCTX-M-3 gene in 3 strains, blaCTX-M-14 gene in 18 strains, blaTEM-1 gene in 16 strains, blaSHV-12 gene in 17 strains, and rmtB in 5 strains. All the 18 CR-HMKP strains carried virulence-associated genes, including rmpA (88.9％, 16/18), magA (5.6％, 1/18), iroN (83.3％, 15/18), aerobactin (27.8％, 5/18), rmpA2 (66.7％, 12/18) and mrkD (100％, 18/18). Three sequence types (STs) were identified by MLST, including ST11 (15 strains), ST86 (2 strains), and ST412 (1 strain). PFGE resulted in three major PFGE clusters, of which cluster A corresponds to ST1 isolates, and cluster B corresponds to ST86 isolates, and cluster C corresponds to ST412 isolates. All the blaKPC-2- positive strains belonged to ST11. Plasmid conjugation was successful in 5 (27.8％) of the 18 CR-HMKP isolates. Conclusions wzi64-K14.64 is the predominant capsule serotype of the CR-HMKP strains in this hospital. KPC-2 gene conjugationmay contribute to the emergence of CR-HMKP isolates. In addition, CRHMKP strain may be the highly prevalent ST11, and highly virulent CPS serotypes harboring K1/K2.

Objective To investigate the effects of morroniside on the expression of vascular endothelial growth factor (VEGF) and fi-broblast growth factor-2 (FGF-2) in rat cortex after focal cerebral ischemia-reperfusion. Methods 30 male Sprague-Dawley rats were ran-domly divided into sham group, model group, morroniside-low group (30 mg/kg), morroniside-middle group (90 mg/kg) and morroni-side-high group (270 mg/kg). Middle cerebral arteries of rats were occluded for 30 minutes with Longa's method and re-perfused. The ex-pression of VEGF and FGF-2 in the ischemic ipsilateral cortex was detected with Western blotting 7 days after reperfusion. Results The ex-pression of both VEGF and FGF-2 increased in the ischemic ipsilateral cortexin in all the ischemic groups compared with the sham group (P<0.05). The expression of VEGF further increased in a dose-dependent manner in all the morroniside groups compared with that of model group (P<0.05), and the expression of FGF-2 increased in the morroniside-high group (P<0.001). Conclusion Morroniside could increase the expression of VEGF and FGF-2 after ischemia-reperfusion, which might promote angiogenesis.