Objective To compare the effect of airway clearance system (VestTM system) and traditional person knocking method on expectoration and pulmonary infection prevention in elderly patients undergone abdominal operation. Methods 60 cases of elderly patients undergone abdominal operation were randomly divided into observation group (n = 30) and control group (n = 30). The control group received person knocking method to promote expectoration while the observation group was treated with VestTM system.Clinical effect was compared between the groups. Results Sputum volume on day 1 of the observation group [(25.32 ± 2.12) ml/d] was higher than that of the control group [(14.20 ± 1.58) ml/d], and the difference was statistically significant (t = 23.04, P ＜ 0.05). There were also significant differences of sputum volume on day 2, day 3 and day4 (P＜0.05). 6 cases of cut pain, (4.2±0.2) scores of VAS and3 cases of difficulty in position cooperation were reported in the observation group, while the control group reported 21 cases,(7.2±0.3) scores and 16 cases, respectively, the differences were statistically significant (x2 = 13.20,11.09, respectively, t = 45. 57, P ＜ 0.05). There was no significant difference regarding hemodynamics and drainage volume (P ＞ 0.05). Conclusions Compared with traditional person knocking method, the VestTM system has satisfying effect of expectoration and popularity with patients, and can effectively prevent pulmonary infection in elderly patients undergone abdominal operation.

As a new hypnotic, zopiclone is widely used in clinical treatment. There are many methods for determination of zopiclone, including spectrophotometry, chromatography and chromatography mass spectrum, etc. Present paper reviews different kinds of biological samples associated with zopiclone, ex-traction and purification methods, and determination and analysis methods, which aims to provide refer-ences for the relevant research and practice.

OBJECTIVETo identify the putative specific localization signal sequence of tumor metastasis suppressor gene-1 (TMSG-1) and to explore the mechanism of subcellular localization of TMSG-1 protein.

METHODSVectors expressing green fluorescence protein (GFP) tagged different TMSG-1 fragments were generated and transfected into human embryo kidney 293 (HEK293) cells. The expression of those fusion proteins was detected by Western blotting and their subcellular localizations were observed by laser confocal microscope.

RESULTSGFP was fused with the native TMSG-1(aa1-380) or different fragments including T1 (aa1-70), T2 (aa1-128), T3 (aa129-380), T4 (aa71-128), T5 (aa71-179) and T6 (aa71-380). Anti-GFP Western blotting showed that these fusion proteins were successfully expressed. Under laser confocal microscope, GFP fused with fragment T4 (aa71-128) localized mainly in the nucleolus; GFP fused with fragment T6 (aa71-380) localized diffusely in the nucleus; while other fusion proteins with TMSG-1 (aa1-380) or fragment T1 (aa1-70), T2 (aa1-128), T3 (aa129-380) and T5 (aa71-179) localized in the cytoplasm. Fragment T4(Δ119-128) was generated from T4 with deletion of 10 amino acid of the C terminal. GFP fused with fragment T4(Δ119-128) remained in the nucleus, but no longer in the nucleolus.

CONCLUSIONSThere is a nucleolar localization signal (aa119-128 RRRRNQDRPS) within TMSG-1. This finding may have laid the foundation for further investigations into subcellular localization and function of TMSG-1.

Amino Acid Sequence ; Blotting, Western ; Cell Nucleolus ; metabolism ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Green Fluorescent Proteins ; metabolism ; HEK293 Cells ; Humans ; Membrane Proteins ; genetics ; metabolism ; Microscopy, Confocal ; Nuclear Localization Signals ; Plasmids ; Recombinant Fusion Proteins ; metabolism ; Sphingosine N-Acyltransferase ; genetics ; metabolism ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism

OBJECTIVETo investigate the regulatory mechanism of the transcription of tumor metastasis suppressor gene TMSG-1.

METHODSLuciferase reporter assay and site-directed mutagenesis were used to analyze the regulatory region of TMSG-1. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) were carried out to verify the interaction of KLF6 and Sp1 with the regulatory region of TMSG-1. Co-immunoprecipitation (CoIP) was performed to analyze the interaction between KLF6 and Sp1. TMSG-1 and wt-KLF6 mRNA expressions in cells with different metastatic capacities were quantitated by real-time PCR. Cell invasive capability was determined by Matrigel invasion assay.

RESULTSA 63 bp inducible regulatory region (+59 bp - +123 bp) in exon 1 was identified by luciferase assay using reporter plasmids with a series of TMSG-1 regulatory region deletions. Mutations in KLF6/Sp1 binding sites of this region resulted in a decrease of luciferase activity, while cotransfection with KLF6 or Sp1 expressing plasmids led to a remarkable increase of luciferase activity. EMSA and ChIP demonstrated that KLF6 as well as Sp1 interacted with this region. CoIP also indicated a possible interaction between KLF6 and Sp1 proteins. In the highly metastatic cell sublines, a low level of wild type KLF6 was associated synchronously with a low TMSG-1 level. Prostate carcinoma cells overexpressing KLF6 exhibited a higher TMSG-1 level and a lower invasive capability.

CONCLUSIONSTranscription factor complex of KLF6 and Sp1 may participate in the inducible transcriptional regulation of TMSG-1, and a decreased wild type KLF6 expression is likely associated with a low TMSG-1 level in the highly metastatic cell sublines.

Binding Sites ; genetics ; Cell Line, Tumor ; Electrophoretic Mobility Shift Assay ; Humans ; Immunoprecipitation ; Kruppel-Like Factor 6 ; Kruppel-Like Transcription Factors ; genetics ; metabolism ; Lung Neoplasms ; metabolism ; pathology ; Male ; Membrane Proteins ; genetics ; metabolism ; Mutagenesis, Site-Directed ; Mutation ; Neoplasm Invasiveness ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Sp1 Transcription Factor ; genetics ; metabolism ; Sphingosine N-Acyltransferase ; genetics ; metabolism ; Transcriptional Activation ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism

Objective To explore the epidemics of influenza viruses in Guangzhou area from 1998 to 2003. Methods The specimens for viral isolation were taken with swabs from children's throats and the material was inoculated into the MDCK cells and were incubated at 33 ℃ The supernatant of MDCK cells culture was tested with hemagglutination test. Results Influenza viruses were isolated from 264 of 3444 children; total positive rate of influenza virus isolation was 7.6%. The positive rate of influenza viruses was 16.8% in 1998; the prevailing strain of influenza viruses was H3N2. The influenza viruses isolation rate was 7.4% in 1999;the positive rate was 8.4% ; HIN1 occurred in 2000, the positive rate was 3.8%. H3N2 did not occur in 2001; the positive rate was 7.3% ; influenza B viruses was the prevailing strain in 2002; the positive rate was 1.7% in 2003. Influenza B viruses was Yamagata like strain from 1998 to 2001, Victoria like strain from 2002 to 2003. H9N2 avian influenza virus was isolated from a child. Conclusions Influenza was prevalent in Guangzhou in 1998, but not prevalent from 1999 to 2003. Most of influenza B viruses were Yamagata strain. There were cases avian influenza caused by H9N2 in 1999.

AIM:To explore the effect of microRNA-221 (miR-221) on the proliferation of lung cancer A549 cells, and to investigate its mechanism .METHODS: The A549 cells were transfected with miR-221 mimics by Lipo-fectamine 2000.The expression of miR-221 was detected by RT-qPCR.The expression of PTEN at mRNA and protein le-vels was detected by RT-qPCR and Western blot , respectively .The cell proliferation was examined by CCK-8 assay and colony formation assay.The 3'-UTR of PTEN was cloned into luciferase reporter vector and its enzymatic activity was detec-ted to verify whether miR-221 targeted to PTEN.RESULTS:The expression level of miR-221 in the A549 cells was sig-nificantly increased after transfection with miR-221 mimics as compared with negative control group and blank group ( P<0.01).The mRNA and protein levels of PTEN were significantly down-regulated compared with control group and blank group ( P <0.05 ) .In addition , miR-221 over-expression significantly promoted the proliferation of A 549 cells ( P <0.05).Moreover, miR-221 inhibited the enzymatic activity of luciferase reporter vector of PTEN.CONCLUSION:Over-expression of miR-221 significantly promotes the proliferation ability of human lung cancer A 549 cells by down-regulation of PTEN.

OBJECTIVE: To analyze the clinical pathologic characteristics of cervical intraepithelial neoplasia grade III (CINIII ) and to explore optimal surgery for CINIII patients. METHODS: The clinical pathologic characteristics, surgical treatments, prognosis and history of 383 CINIII patients, who hospitalized from August 2005 to December 2010, were reviewed and analyzed. Among the patients, 213 (55.6%) received cold-knife conization surgery and 170 (44.4%) received ordinary electric knife conization surgery. RESULTS: There was no significant statistic difference between cold-knife conization group and ordinary electric-knife conization group on the level of clearance of the pathologic tissues and the cervical cone diameter and cone high. Intraoperative blood loss was (13.1±5.2) mL and (25.5±17.2) mL. Bleeding of electric knife conization group, compared with that of the cold knife conization group, decreased by nearly 50%. The difference between the 2 groups was significant (P<0.01). Pathological examination after conization operation indicated that 350 out of the 383 patients didn't show pathological upgrade while 33 patients showed pathological development, among which 21 were diagnosed with invasive cervical cancer at Ia1 clincal stage, 7 atIa2 clincal stage and 5 atIb1 clincal stage. In 3 cases (14.3%) Ia1 cervical cancer patients, fertility requirements and negative margins with cervical conization were closely followed up, and one patient (4.8%) with positive margin and fertility requirements had re-conecut. The remaining 17 (80.9%) had resected the uterus outside the fascia (or plus attachments) . All the 12 patients with invasive cervical cancer at Ia2 orIb1 clinical stage received radical hysterectomy. No tumor recurrence was observed in the 383 patients. CONCLUSION Treatment optimazation of CINIII patients should be based on clinical pathological diagnosis and individual requirements. Both cervical conization surgery and total hysterectomy have been proved safe and practical for CINIII patients.
Adult ; Aged ; Cervical Intraepithelial Neoplasia ; pathology ; surgery ; Conization ; methods ; Female ; Humans ; Hysterectomy ; Middle Aged ; Neoplasm Grading ; Uterine Cervical Neoplasms ; pathology ; surgery

Objective To compare the ability and influence factors of three platelet function tests:light transmission aggregometry (LTA),thrombelastograph (TEG) and VerifyNow.Methods A total of 174 patients underwent percutaneous coronary intervention (PCI) and platelet function test were enrolled.Platelet function were tested by LTA,TEG and VerifyNow.Results Patients were found to be high platelet reactivity (HPR) by LTA (56.64％,64 cases/113 cases),TEG(60.38％,32 cases/52 cases),VerifyNow (17.24％,15 cases /87 cases).There were no significant differences observed between TEG and LTA (P ＞0.05).VerifyNow identified lower HPR proportion compared with TEG and LTA (all P ＜ 0.001).Conclusion Compared with LTA and TEG,VerifyNow identified lower HPR proportion and may overestimate the effectiveness of clopidogrel.We will follow the patients to study the predictive capability of three tests.

Objective To investigate the differences in distribution of platelet endothelial aggregation receptor 1 (PEAR1) between Chinese cardiovascular patients and healthy population. Methods This study used method of Sequenom Mass ARRY to detect differences in distribution of PEAR1 gene between Chinese cardiovascular patients and healthy population. Results A total of 203 patients with cardiovascular disease and 106 healthy volunteer were enrolled. Nineteen sites were detected and all of them are variants in PEAR1 locus. There is no statistical difference of distribution between patients and healthy volunteers(P > 0. 05). Conclusion PEAR1 gene have multiple polymorphic sites in Chinese people.

PURPOSE: CO₂ leakage along the trocar (chimney effect) has been proposed to be an important factor underlying port-site metastasis after laparoscopic surgery. This study aimed to test this hypothesis by comparing the incidence of port-site metastasis between B-ultrasound-guided and laparoscopically-assisted hyperthermic intraperitoneal perfusion chemotherapy (HIPPC). MATERIALS AND METHODS: Sixty-two patients with malignant ascites induced by gastrointestinal or ovarian cancer were divided into two groups to receive either B-ultrasound-guided or laparoscopically-assisted HIPPC. Clinical efficacy was assessed from the objective remission rate (ORR), the Karnofsky Performance Status (KPS) score, and overall survival. The incidence of port-site metastasis was compared between the two groups. RESULTS: Patients in the B-ultrasound (n=32) and laparoscopy (n=30) groups were comparable in terms of age, sex, primary disease type, volume of ascites, and free cancer cell (FCC)-positive ascites. After HIPPC, there were no significant differences between the B-ultrasound and laparoscopy groups in the KPS score change, ORR, and median survival time. The incidence of port-site metastasis after HIPPC was not significantly different between the B-ultrasound (3 of 32, 9.36%) and laparoscopy (3 of 30, 10%) groups, but significantly different among pancreatic, gastric, ovarian, and colorectal cancer (33.33, 15.79, 10.00, and 0.00%, p<0.001). CONCLUSION: The chimney effect may not be the key reason for port-site metastasis after laparoscopy. Other factors may play a role, including the local microenvironment at the trocar site and the delivery of viable FCCs (from the tumor or malignant ascites) to the trauma site during laparoscopic surgery.
Ascites ; Colorectal Neoplasms ; Drug Therapy* ; Humans ; Incidence ; Karnofsky Performance Status ; Laparoscopy ; Neoplasm Metastasis* ; Ovarian Neoplasms ; Perfusion* ; Surgical Instruments ; Treatment Outcome