Objective:To study the influence of Wnt5a gene silence on the proliferation,migration and invasion of lung squamous carcinoma cells.Methods:A recombinant plasmid pHI-siRNA~(Wnt5a-)was constructed and used to deliver small interference RNA (siRNA)targeting Wnt5a in SK-MES-1 cells;the transfected cells were screened to establish a stable transgenic cell line.MTT,cell cycle and Transwell assays were employed to evaluate the effect of Wnt5a gene silence on the proliferation,migration and invasion of lung squamous carcinoma cells.Results:Western blotting assay revealed that Wnt5a was lowly expressed in SK-MES-1~(Wnt5a-)(13.6%).The proliferation index(PI)of transgenic cell line was slightly lower than that of the control cell line([28.3?3.8]% vs[30.5?5.2]%). The migration and invasion capabilities of SK-MES-1~(Wnt5a-)cells were(47.3?9.2)% and(39.7?11.7)% of the control cells, respectively.Conclusion:Low Wnt5a expression can significantly inhibit the migration and invasion capabilities of SK-MES-1 cells, indicating that Wnt5a might be a potential target for gene therapy of lung squamous carcinoma.

Objective To study the role of the outer membrane protein Rmp of Neisseria gonor-rhoeae strain in immunosuppression and the strategy of eliminating it .Methods The rmp gene of Neisseria gonorrhoeae strain was amplified by PCR and inserted into pMD 19-T vector .The recombinant vector pMD 19△rmp∷Kan containing Kan and the 5′-and 3′-flanking regions of rmp (△rmp∷Kan) was constructed by replacing 200 nucleotide residues of pMD 19-rmp with kanamycin resistance gene Kan and transformed into Neisseria gonorrhoeae WHO-A strain.PCR and Western blot assay were used to screen and identify the re-combinant mutant strains that could not express Rmp .Mice were immunized with mutant strains and bacteri-cidal activities of the immune sera were detected by antibody-mediated complement-dependent cytotoxicity assay.Results The mutant strains that could not encode Rmp were successfully constructed .Antibodies in-duced by mutant strains showed stronger bactericidal activity against Neisseria gonorrhoeae in comparison with those induced by wild strains .Conclusion The recombinant Neisseria gonorrhoeae strain with rmp gene de-letion might eliminate the immunosuppressive effects of Rmp expressed in wild gonococcal strains , which provides a reference for further development of novel live attenuated whole-cell vaccines of Neisseria gonor-rhoeae.

Objective To discuss the treatment of renal or adrenal tumor with cancer thrombus in the inferior vena cava. Methods From Jan 1984 to Apr 2008,29 cases of renal or adrenal malignancy with thrombosis involving the inferior vena cava underwent treatment.The diagnoses were confirmed by Doppler uhrasonography,CT and MRI.In the 29 surgical patiens the tumor thrombus was level I in 7,level Ⅱ in 10,level Ⅲ in 8 and levelⅣin 4.According to TNM classification,23 cases were classified to T2N.M.,1 case was T2Nl Mo,1 case was TzNlMl,1 case was T3NoMo,2 case were T3NlMl and 1 case was T3N2Mo.The mean tumor size was 8.7(4.O-16.O)cm in diameter.The mean tumor thrombosis length was 3.2(2.5-4.0)cm in level I,5.3(4.5-6.0)cm in level Ⅱ,8.2(6.5-9.O)cm in levelⅢand 15.1(12.0-18.5)cm in level IV. Results The operation was performed succesgfullv in 29 patients Patholocieal examination showed that 18 cases of clear cellcarcinoma,3 cases sarcomatoid carcinoma,2 cases renal papillary adenocarcinoma,1 case renal cell carcinoma (undifferentiated),1 case granule carcinoma,3 cases adrenocortical carcinoma and 1 case metastatic malignant melanoma of adrenal gland.Of 29 patients,3 were out of contact.Twenty-six patients were followed up for 35(0-62)months after treatment,3-and 5-year survival rates were 15/26 and 11/26.Three-year survival rates for stage T2 and T3 were 14/22 and 1/4.Five-year survival rates for stage T2 and T3 were 10/22 and 1/4.Three-year survival rates for level I、Ⅱ、Ⅲ andⅣ were 4/6,5/8,5/8 and 1/4.Five-year survival rates for level I,Ⅱ、Ⅲ andⅣ were 3/6,4/8,3/8 and 1/4.Three-year survival rates for a tumor thrombus in the below or above diaphragm were 14/22 versus 1/4,5-year survival rates were 10/22 versus 1/4.Three-year and 5-year survival rates for the patients without distant metastases and lymph node involvement were 12/18 and 9/18.Three-year and 5-year surviral rates for the patients with distant metastases and lymph node involvement were 3/8 and 2/8.The 3 surgical patients with metastatic disease died at 6,10,22 months. Conclusions Surgical treatment could be the preferred approach for the patients of renal or adrenal tumor with cancer thrombus in the inferior vena cava without distant metastases and lymph node involvement.It could improve the quality of life and may prolong survival.

Objective:To explore the related risk factors of antepartum depression and anxiety,and to provide the theoretical support for early screening,clinical intervention and prevention of antepartum depression and anxiety.Methods:At the time of 28 weeks of prenatal examination,the Edinburgh Postnatal Depression Scale (EP-DS),Self-Rating Anxiety Scale (SAS)were used to survey 2112 pregnant women who were selected from Maternal and Child Health Hospital of Kunming City.Non conditional logistic regression analysis was used to explore the re-lated risk factors of prenatal depression and anxiety.Results:At the 28 weeks of pregnancy,the detection rate of de-pression symptoms and anxiety symptoms were 25.4% and 6.6%.Logistic regression analysis showed that the risk factors for perinatal depression symptoms were younger age (OR =0.80,95%CI:0.68 -0.94),not stick to work during pregnancy (OR =1.18,95%CI:1.02 -1.36),not satisfied with the living environment (OR =1.50,95%CI:1.23 -1.83),expectations for boys (OR =0.86,95%CI:0.77 -0.96),not only daughter (OR =1.37,95%CI:1.06 -1.76),unplanned pregnancy (OR =1.38,95%CI:1.10 -1.72).The risk factors for perinatal anxiety symp-toms were being not satisfied with the living environment (OR =1.64,95%CI:1.19 -2.26),not harmonious with her husband (OR =2.01,95%CI:1.20 -3.37),unplanned pregnancy (OR =1.50,95%CI:1.05 -2.14).Conclu-sion:It suggests that the pregnant women with younger age,less working during pregnancy,being not satisfied with the living environment,having more expectations for boys,being not only daughter,with unplanned pregnancy are more likely to suffer from antepartum depression;those being not satisfied with living environment,having harmoni-ous relation with her husband,and with unplanned pregnancy are more likely to suffer from antepartum anxiety.

OBJECTIVETo investigate the intervention of Naoxintong and mecobalamin on electrophysiological changes in diabetic peripheral neuropathy (DPN) of different Chinese medicine (CM) syndrome types.

METHODSAccording to syndrome differentiation, 180 patients with DPN were classified as five syndrome types. And they were treated with Naoxintong (Group A), mecobalamin (Group B), and Naoxintong + mecobalamin (Group C). Four weeks was taken as one therapeutic course, and totally three courses. Their efficacies were assessed using clinical scoring, electrophysiological examinations, and ultrasonic examinations of the blood vessel inner diameter.

RESULTS(1) The motor nerve conduction velocity was obviously slowed down in the Gan-Shen deficiency syndrome (P<0.01). F-wave latency was obviously prolonged in the Gan-Shen deficiency syndrome and yang deficiency blood stasis syndrome (P<0.01). The skin sympathetic reflex latency was obviously prolonged in the qi deficiency blood stasis syndrome and phlegm stagnation collateral obstruction syndrome (P<0.01). (2) Statistical difference existed in the three groups of qi deficiency blood stasis syndrome (chi2 = 7.112, P<0.05) and Gan-Shen deficiency syndrome (chi2 =6.667, P<0.05). Of them, the total effective rate of qi deficiency blood stasis syndrome was 87.5% and the markedly effective rate 43.8% in Group A (P<0.05). The total effective rate of Gan-Shen deficiency syndrome was 100.0% and the markedly effective rate 50.0% in Group B (P<0.05). The total effective rate of qi deficiency blood stasis syndrome, yin deficiency blood stasis syndrome, phlegm stagnation collateral obstruction syndrome, yang deficiency blood stasis syndrome, and Gan-Shen deficiency syndrome was respectively 92.9%, 83.3%, 81.8%, 81.8%, and 75.0% in Group C. (3) Naoxintong and mecobalamin had some improvement of motor and sensory conduction of each CM syndrome type (P<0.05). Mecobalamin showed obvious effect on the skin sympathetic reflection (P<0.05). The nerve electrophysiological index of each syndrome types as well as the diameter of arteriae tibialis anterior could be improved in Group C (P<0.05).

CONCLUSIONSNaoxintong gained better effect in treatment of DPN patients of qi deficiency blood stasis syndrome by syndrome typing. Naoxintong combined with mecobalamin could be helpful for ameliorating DPN patients of each syndrome.

Adult ; Diabetic Neuropathies ; diagnosis ; drug therapy ; physiopathology ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Electrophysiological Phenomena ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Peripheral Nervous System Diseases ; diagnosis ; drug therapy ; physiopathology ; Phytotherapy ; Vitamin B 12 ; analogs & derivatives ; therapeutic use

AIMTo prepare solid lipid nanoparticles by microemulsion technique.

METHODSStearic acid was used as the oil phase, lecithin as surfactant, alcohol as cosurfactant and distilled water as the aqueous phase. Microemulsion was prepared by mixing the above component in proper ratio. The corresponding pseudoternary phase diagram monitored Microemulsion formation field of different lecithin/alcohol. Solid lipid nanoparticles (SLN) were prepared by dispersing warm microemulsion in cold water under magnetic stirring. Then appropriate microemulsions that can contain more water phase and suitable oil phase were selected to prepare SLN. The influence of formulation, process variables on the preparation and quality of SLN were studied. Based on the investigation of single factors, orthogonal design was used to optimize SLN formulation and preparation process, and more, the reproducibility of the optimized results were studied.

RESULTSThe results showed that the device temperature (Ti), water temperature (Tw), and delivery rate (Rd) were the key factors that influence the preparation process of SLN, and Tw was extremely important. The ratio of microemulsion formulation, the ratio of microemulsion and distilled water had also influence on its quality.

CONCLUSIONMicroemulsion technique can be used to prepare solid lipid nanoparticles.

Alcohols ; Drug Carriers ; Emulsions ; Lipids ; chemical synthesis ; chemistry ; Nanotechnology ; Particle Size ; Phosphatidylcholines ; Solubility ; Technology, Pharmaceutical ; methods

Objective To investigate correlation between aspirin resistance(AR) and inflammatory factors. Methods One hundred and ten patients with coronary heart disease took aspirin 0.1 mg/d for 14 days.It was detected platelet aggregation function induced with adenosine disphosphate (ADP) and arachidonic acid (AA), and investigated correlation between AR and inflammatory factors. Interleukin-1? (IL-1?),interleukin-6 (IL-6) and high sensitive C-reaction protein (hs-CRP) levels. Results IL-6 level of patients with AR was significantly higher than that of aspirin sensitive (AS) patients. The other two index were not different between the two groups. Conclusion IL-6 levels could be used as predictor.

OBJECTIVE: To evaluate the application value of Toronto clinical scoring system (TCSS) and its grading of neuropathy for diabetic peripheral neuropathy (DPN), and to explore the relationship between TCSS grading of neuropathy and the grading of diabetic nephropathy and diabetic retinopathy. METHODS: A total of 209 patients of Type 2 diabtes (T2DM) underwent TCSS. Taking electrophysiological examination as a gold standard for diagnosing DPN, We compared the results of TCSS score > or = 6 with electrophysiological examination, and tried to select the optimal cut-off points of TCSS. RESULTS: The corresponding accuracy, sensitivity, and specificity of TCSS score > or = 6 were 76.6%, 77.2%, and 75.6%, respectively.The Youden index and Kappa were 0.53 and 0.52, which implied TCSS score > or = 6 had a moderate consistency with electrophysiological examination. There was a linear positive correlation between TCSS grading of neuropathy and the grading of diabetic nephropathy and diabetic retinopathy (P<0.05). The optimal cut-off point was 5 or 6 among these patients. CONCLUSION TCSS is reliable in diagnosing DPN and its grading of neuropathy has clinical value.
Adult ; Aged ; Diabetes Mellitus, Type 2 ; complications ; Diabetic Neuropathies ; diagnosis ; physiopathology ; Electrophysiology ; Female ; Humans ; Male ; Middle Aged ; Neurologic Examination ; methods

OBJECTIVETo study mRNA expression of immune-related genes (Foxp3, GATA3, CTLA4 and T-bet) in peripheral blood of the patients with allergic dermatitis induced by trichloroethylene (TCE).

METHODSThe peripheral blood samples were collected from 8 healthy workers (control group) and 8 patients with allergic dermatitis induced by TCE (case group). Real-time quantitative PCR was applied to detect mRNA expression of immune-related genes (Foxp3, GATA3, CTLA4, T-bet).

RESULTSThe mRNA expression levels of Foxp3, GATA3 and CTLA4 genes increased by 115%, 97% and 241% in case group, as compared with control group (P < 0.01). The mRNA expression level of T-bet gene decreased by 47% in case group, as compared with control group (P < 0.01).

CONCLUSIONThe mRNA expression levels of some immune-related genes changed in patients with allergic dermatitis induced by TCE, those genes may play an important role in TCE-induced allergy.

Adult ; CTLA-4 Antigen ; metabolism ; Case-Control Studies ; Dermatitis, Occupational ; genetics ; immunology ; Female ; Forkhead Transcription Factors ; metabolism ; GATA3 Transcription Factor ; metabolism ; Gene Expression ; Humans ; Male ; RNA, Messenger ; genetics ; T-Box Domain Proteins ; metabolism ; Trichloroethylene ; Young Adult

OBJECTIVETo prepare cytochrome (CYP)2E1-silenced hepatocytes by lentivirus-mediated RNA interference technology and to investigate the hepatotoxicity of trichloroethylene (TCE) in CYP2E1-silenced hepatocytes.

METHODSShort hairpin RNA fragments were designed and synthesized and were then ligated into the lentiviral vector; single colonies were screened; the plasmid was extracted after PCR and sequence identification and then transferred into L02 hepatocytes; the CYP2E1-silenced hepatocytes were selected; real-time quantitative PCR and Western blot were used to evaluate the interference effects. The obtained CYP2E1-silenced hepatocytes, as well as normal L02 hepatocytes, were treated with TCE (0, 0.25, 0.50, 1.00, 2.00, and 4.00 mmol/L). The cell viability and half maximal inhibitory concentration (IC50) of TCE were measured; the apoptotic rate of cells was measured by flow cytometry; the mRNA expression levels of apoptosis genes and oncogenes were measured by real-time quantitative PCR.

RESULTSThe IC50s of TCE for L02 hepatocytes and CYP2E1-silenced hepatocytes were 15.1 mmol/L and 23.6 mmol/L, respectively. The apoptotic rate increased as the dose of TCE rose in the two types of cells; the CYP2E1-silenced hepatocytes hada significantly lower apoptotic rate than L02 hepatocytes when they were exposed to 2.0 and 4.0 mmol/L TCE (P < 0.05 or P < 0.01). The mRNA expression level of bcl-2 (anti-apoptosis gene) in CYP2E1-silenced hepatocytes was 15% ∼ 60% higher than that in L02 hepatocytes (P < 0.01), while the mRNA expression levels of caspase-3 and caspase-9 (apoptosis genes) in CYP2E1-silenced hepatocytes were 30% ∼ 60% lower than those in L02 hepatocytes (P < 0.01). The mRNA expression level of p53 (cancer suppressor gene) in CYP2E1-silenced hepatocytes was 81 - 278% higher than that in L02 hepatocytes (P < 0.01), while the mRNA expression levels of c-fos and k-ras (oncogenes) in CYP2E1-silenced hepatocytes were 20-68% lower than those in L02 hepatocytes (P < 0.01).

CONCLUSIONCYP2E1-silenced cells can be successfully prepared by lentivirus-mediated RNA interference technology. Silencing CYP2E1 gene can reduce the hepatotoxicity of TCE and inhibit the expression of some apoptosis genes and oncogenes, suggesting that CYP2E1 gene plays an important role in TCE metabolism and is related to the hepatotoxicity of TCE.

Apoptosis ; drug effects ; genetics ; Cell Line ; Cell Survival ; drug effects ; genetics ; Cytochrome P-450 CYP2E1 ; genetics ; metabolism ; Genetic Vectors ; Hepatocytes ; drug effects ; metabolism ; Humans ; Lentivirus ; genetics ; RNA Interference ; Trichloroethylene ; toxicity