OBJECTIVE:To investigate the therapeutic efficacy of recombination human growth hormone(rhGH)on the patients with chronic obstructive pulmonary disease(COPD)complicated with spontaneous pneumothorax(SP).METHODS:58 patients with COPD complicated with SP were randomly divided into two groups:29 cases in the control group received conventional therapy including anti-infection therapy,oxygen therapy and nutritional support,and another 29 in the trial group received low dose of rhGH(0.1 U?kg-1?d-1)hypo quaque nocte for 7 days started from the second day after admission in addition to the conventional therapy as employed for control group.The time of chest tube stay and the length of hospital stay were compared between the two groups.The body mass index(BMI)and serum levels of growth hormone(GH),insulin-like growth factor-1(IGF-1)and albumin(ALB)were measured in all cases before and after treatment.RESULTS:The time of chest tube stay and the length of hospital stay in the trial group vs.the control group were(5.5?1.8)d vs.(9.3?2.2)d(P0.05);however,after treatment,all indexes(BMI,ALB,GH and IGF-1)were higher in the trial group than in the control group(P

Objective To investigate the adjunctive therapeutic effect of recombination growth hormone(rhGH) on the elderly patients with pulmonary infection.Methods 53 elderly patients with pulmonary infection were randomly divided into rhGH group(n=25) and control group(n=28).The rhGH group were received hypodermic introl group were received standard therapy.Clinical efficacy were compared between two groups.The level of albumin (ALB),growth hormone (GH),insulin-llke growth factor-1 (IGF-1),leptin (LP) were measured and the body mass index(BMI) was observed in all cases before and after treatment.Results The overall efficacy of rhGH group and control group was 88.0% versus 60.7%.The results showed significant statistical difference between both groups(P＜0.05).After treating with rhGH for 10 days,the level of BMI,ALB,GH,IGF-1 and LP were increased in the rhGH group,which were(26.1±4.1)kg/m2;(38.4 ±6.6)g/L;(7.0 +0.9) μg/L;(27.3 ±6.1)μg/L;(6.9 ± 1.1)μg/Lrespectively.And the level of BMI,ALB,GH,IGF-1 and LP were (21.8 ± 3.4 ) kg/m2 ;( 29.5 ± 5.1 ) g/L;( 4.0 ±0.4) μg/L;( 22.0±3.8 )μg/L;( 3.8±0.8 )μg/L in the control group after treatment.The results showed significant statistical difference between both groups(P＜0.05).Conclusion There was obvious adjunctive therapeutic effect of rhGH to the elderly patients with pulmonary infection by improving the nutritional state of these patients.

Objective To establish a stable cell line expressing transmembrane form of human blood group A antigen mimotope vaccine by transfecting malignant melanoma cell line B16, and to detect the cytotoxicity of the vaccine against melanoma cells. Methods Cultured B16 cells were classified into 4 groups, i.e.,P/F-M-pIRES group [transfected with the recombinant plasmid mimotope peptide/Fas-macrophage inflammatory protein (Mip)-pIRES], P/F-pIRES group (transfected with the recombinant plasmid mimotope peptide/FaspIRES), M-pIRES group (transfected with the recombinant plasmid Mip-pIRES), and pIRES group (transfected with the empty plasmid pIRES). B 16 cells were transfected through Lipofectamine 2000. Subsequently, RT-PCR and Western blotting were performed to detect the mRNA and protein expressions of the mimotope peptide/Fas fusion gene and Mip3β in transfected B16 cells. Cell counting kit-8 (CCK-8) was used to evaluate the vaccinemediated complement dependent cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC)against B16 cells. Results RT-PCR yielded specific DNA fragments with expected size. Western blotting revealed the anti-A antibody-binding activity of the recombinant mimotope peptide/Fas fusion protein. Factor analysis indicated significant differences in CDC (F = 244.522, P ＜ 0.01 ) and ADCC (F = 71.593, P ＜ 0.01 )against B16 cells between the 4 groups. Group comparisons demonstrated more intense CDC and ADCC in P/FM-pIRES and P/F-pIRES groups compared with M-plRES and pIRES groups, stronger ADCC in P/F-M-pIRES group in comparison with P/F-pIRES group (F = 15.42, P ＜ 0.05), but no significant difference in CDC was observed between M-pIRES and pIRES group. Conclusions The transmembrane form of human blood group A antigen mimotope vaccine could be stably expressed in B16 cells, and mediate ADCC and CDC against B16 cells in vitro.

Objective To investigate the antitumour effects of transfected gut-enriched Krüppellike factor(GKLF) on human gastric carcinoma cell line SGC-7901 in vitro and in vivo. Methods The expression of GKLF mRNA and protein in human gastric carcinoma cell line SGC-7901 were detected before and after transfection by real-time fluorescence quantitative PCR and Western blot,respectively. Proliferation and invasion in SGC-7901 were measured respectively by MTT assay, flow cytometry, colony formation assay and cell invasion assay after transfected with GKLF. The growth of xenograft was observed, the microvessel density(MVD) of xenograft tissue was determined by immunohistochemistry. Results The GKLF mRNA and protein in SGC-7901 were overexpressed after transfected with GKLF(P＜0.05). The proliferative speed of SGC7901-pcDNA3.1-GKLF group was markedly lower than that of SGC-7901 and SGC7901-pcDNA3.1 groups (P＜0.05). Transfected with GKLF caused part of the G0/G1 arrest, decreased clone formation rate and the invasion ability (P＜0.05). The growth speed of xenograft in SGC7901-pcDNA3.1-GKLF group was lower, the weight and MVD of xenograft tissue in SGC7901-pcDNA3. 1-GKLF group were less (P＜ 0. 05).Conclusion Transfected with GKLF maysuppress proliferation and invasion in human gastric carcinoma cell line SGC-7901, inhibit the growth and the angiogenesis of xenograft in nude mice.

Objective : To estimate the prevalence and risk factors of suicide ideation,suicide planning and suicide attempts among middle school students and to provide reference for suicide intervention. Methods : The students of grade 7 to 12 in Ningbo were recruited through multi-stage random sampling method. A self-reported questionnaire survey was conducted to collect the information about suicide ideation,suicide planning and suicide attempts within 12 months. A multivariate logistic regression model was used to analyze the risk factors for the three aspects of suicide among middle school students. Results: A total of 10 729 questionnaires were sent out and 10 726 valid ones were collected,with a validity rate of 99.97%. The prevalence rates of suicide ideation,suicide planning and suicide attempts among middle school students during the last 12 months were 12.93%,4.54% and 5.06%,respectively. The results of multivariate logistic regression analysis indicated that females（OR：1.397-1.575,95%CI：1.178-1.927）,students of grade 7 to 9（OR：1.625-1.824,95%CI：1.323-2.082）,poor health condition（OR：1.160-2.131,95%CI：1.005-2.985）,loneliness（OR：1.574-4.423,95%CI：1.221-5.254 ）,desperation（OR：2.796-3.232,95%CI：2.400-3.990）,anxiety（OR：1.890-2.117,95%CI：1.503-2.496）,less than 8 hours a day of sleep（OR：1.152-1.263,95%CI：1.030-1.594）,smoking（OR：1.476-2.074,95%CI：1.055-3.072）,drinking（OR：1.479-1.863,95%CI：1.271-2.296）,fighting（OR：1.716-1.941,95%CI：1.330-2.422）and school bullying（OR：2.254-3.292,95%CI：1.342-5.277）were common risk factors for suicide ideation,suicide planning and suicide attempts;physical activity（OR：0.597-0.720,95%CI：0.474-0.923）was a common protective factor for suicide ideation,suicide planning and suicide attempts. Conclusion Females,lower grade,poor health condition,loneliness,desperation,anxiety,lack of sleep,smoking,drinking,fighting,school bullying and physical activity were influencing factors for suicidal behaviors.

Objective To investigate the application of colostomy care products in managing the leakage around PTCD drainage tube, and to discuss its clinical effect. Methods A total of 56 patients with malignant obstructive jaundice, who had received PTCD and suffered from postoperative leakage around PTCD drainage tube, were randomly and equally divided into the study group (n=28) and the control group (n=28). The colostomy care powder, the skin protective film and the colostomy bag were used for the patients of the study group, while iodophors, sterile gauze and mupirocin ointment were employed for the patients of the control group. The incidence of irritant dermatitis around PTCD drainage tube, the degree of comfort to the indwelling tube judged by the patient, and the nursing workload for PTCD drainage tube were determined, and the results were compared between the two groups. Results The incidence of irritant dermatitis around PTCD drainage tube in the study group was obviously lower than that in the control group. And the degree of comfort to the indwelling tube judged by the patient in the study group was much higher than that in the control group. The differences between the two groups were statistically significant (P<0.05). Conclusion Combination use of colostomy care powder, skin protective film and colostomy bag can effectively reduce the incidence of irritant dermatitis around PTCD drainage tube and improve the patient’s condition. Therefore, this method is worthy of popularization in clinical practice.

Objective To observe the effects of scutellaria barbata extract (ESB) on proliferation, apoptosis and telomerase activity of human malignant glioma U251 cells in vitro.Methods Different concentrations of ESB (50, 25, 12.5, 6.25, 3.125 and 1.5625 mg/mL) were added into the medium cultured human malignant glioma U251 cells for 24, 48 and 72 h, respectively. And blank control group was also established. MTT assay was employed to detect the proliferation of U251cells. AnnexinV/PI staining and low cytometry (FCM) were used to detect the changes of apoptotic rate.And the telomerase activity of the cells was observed under the examination of telomeric repeat amplification protocol-PCR (TRAP-PCR)-ELISA.Results ESB inhibited the proliferation and induced the apoptosis of U251 cells. Interaction effect was found between the concentration of ESB and the treatment time of ESB by MTT assay (F=59.908, P=0.000); 50 mg/mL ESB for 72 h could most significantly inhibit the proliferation of U251 cells. Interaction effect was found between the concentration of ESB and the treatment time of ESB by AnnexinV/PI staining (F=6.548, P=0.000); 25mg/mL ESB for 72 h could most significantly induce the apoptosis of U251 cells. Interaction effect was also found between the concentration of ESB and the treatment time of ESB by TRAP-PCR-ELISA(F=138.433, P=0.000); the telomerase activity of the cells was the lowest by treatment with 50 mg/mL ESB for 72 h; negative correlation was noted between the telomerase activity of the cells and the apoptosis rate (r=-0.785, P=0.037); so as the telomerase activity of the cells and the inhibition effect (r=-0.278, P=0.042) Conclusion ESB may inhibit the proliferation and induces the apoptosis of U251 cells through down-regulating the telomerase activity.

Objective To determine whether Morphine has the ability to enhance HIV-1 replication in MT2 and Macrophage in vitro and assess the influence of Naloxone on Morphine2s effect.Methods MT2 cells were randomly assigned into 4 groups: (1) Morphine treatment for MT2 group, (2) Morphine+Naloxone co-treatment for MT2 group, (3) Naloxone treatment for MT2 group and (4) MT2 Control;Macrophages were also randomly assigned into 4 groups: (5) Morphine treatment for Macrophage group, (6) Morphine+Naloxone co-treatment for Macrophage group, (7) Naloxone treatment for Macrophage group and (8) Macrophage Control. Group (2), (3), (6) and (7) were pre-treated with 10-8 mol/L Naloxone for 0.5 h, and then group (1) and (2) were treated with 10-12, 10-10 and 10-8 mol/L Morphine for 24 h;group (5) and (6) were disposed of 10-10 mol/L Morphine for 24 h.All 8 groups were added in HIV-1 viral strain with 50% tissue culture infective dose(TCID50).P24 antigen in MT2 cells culture supernatant at day 3, 4, 5 and 6, and in Macrophages culture supernatant at day 4, 6, 8, 10 and 12 after infection were determined with ELISA.Student2s t-test and ANOVA were used to compare the differential expression in different groups, and repeated measures ANOVA was used to compare the increasing or decreasing expression of p24 antigen in morphine treatment groups than that in the control group at different time points.Results On the 3rd day of infection with HIV-1 in MT2 cells, the expression of p24 antigen in 10-12, 10-10 and 10-8mol/L dose of group (1) were (4.44?.30), (5.59?.25) and (4.60?.24) ng/ml respectively, compared to control[(1.93?.05) ng/ml, t= 14.15, 24.74 and 19.14, all P<0.01].On the 4th day, 10-12, 10-10 and 10-8mol/L dose of group (1) resulted in a significant increase of p24 antigen expression [(24.30?.66), (31.73?.17) and (26.02?.37) ng/ml]in culture supernatants compared to control[(8.03?.09) ng/ml, t=10.59, 34.92 and 81.2, all P<0.01].On the 5th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (1) were (56.30?.26), (81.77?.49) and (63.66?.57) ng/ml respectively, compared to control [(15.30?.91) ng/ml, t= 45.83, 43.51 and 30.07, all P<0.01].On the 6th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (1) were (150.70?.97), (243.09?.93) and (173.72?.73) ng/ml respectively, compared to control [(41.01?.84) ng/ml, t= 21.09, 39.02 and 29.55, all P<0.01].The enhanced multiple of p24 antigen expression in three doses of morphine treatment group compared to control increased with HIV-1 infected MT2 cells time, trend analysis of repeated measurements showed statistically significant time effect (F=842.18, P<0.01). On the 4th day of infection with HIV-1 in Macrophage cells, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (5) were (0.68?.15), (0.87?.41) and (0.75?.09) ng/ml respectively, compared to control [(0.60?.01) ng/ml, t= 7.27, 11.06 and 3.02, all P<0.05]. On the 6th day, 10-12, 10-10 and 10-8 mol/L dose of group (5) resulted in a significant increase of p24 antigen expression[(1.64?.57) , (2.07?.12 ) and (1.75?.17) ng/ml]in culture supernatants compared to control [(1.16?.07) ng/ml, t=8.93, 11.3 and 5.45, all P<0.01].On the 8th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (5) were (6.31?.17), (8.81?.34) and (7.19?.11) ng/ml respectively, compared to control [(3.84?.45) ng/ml, t=8.83, 15.11 and 12.42, all P<0.01]. On the 10th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of Morphine treated group were (32.30?7.55), (50.74?7.55) and (39.74?.56) ng/ml respectively, compared to control [(17.55?.86) ng/ml, t= 13.65, 17.84 and 36.69, all P<0.01].The enhanced multiple of p24 antigen expression in three doses of group (5) compared to control increased with HIV-1 infected Macrophage cells time, trend analysis of repeated measurements showed statistically significant time effect (F=135.58, P<0.01).Conclusions Morphine has the ability to enhance HIV-1 replication in MT2 cell and Macrophage. This Morphine-mediated increase of p24 antigen expression can be blocked by Naloxone.

BACKGROUND: Pearl has the tranquilizing effect, and it can be applied in the treatment of hypertension. However, there is little report on the prevention and cure effect and mechanism of hydrolyzed pearl liquid on hypertension. OBJECTIVE: To observe the effect of hydrolysis of Hepu pearl (hydrolyzed Nanzhu fluid) on the biological behavior and secretion of human microvascular endothelial cells. METHODS: Human microvascular endothelial cells were cultured and passaged. There were four groups, and the microvascular endothelial cells were incubated in the 200 μL culture medium containing nothing (control group), and 120, 60 and 30 mg/L hydrolysis of Nanzhu fluid. The cell proliferation and migration was detected by cell conuting kit-8 assay and Transwell assay respectively; the cell cycle distribution was tested by flow cytometry; the cell apoptosis was assayed by TUNEL method; the secretion of nitric oxide and reactive oxygen species was tested by nitrale reduetase and chemical fluorescence method, respectively. RESULTS AND CONCLUSION: Compared with the control group, hydrolysis of Nanzhu fluid significantly promoted the proliferation of microvascular endothelial cells in a manner-dependent manner (P ＜ 0.05), suggesting the optimal concentration was 120 mg/L. Compared with the control group, the percentage of cells in S and G2 phase was significantly increased, and the percentage of cells in the G1 phase was significantly reduced in the hydrolysis of Nanzhu fluid group (P＜0.05), indicating hydrolysis of Nanzhu fluid could promote cell cycle progression. Apoptotic cells with green-stained nucleus were invisible in both groups. The number of cell migration in the hydrolysis of Nanzhu fluid group was significantly more than that in the control group (P ＜ 0.05). Compared with the control group, there was a significant increase in the nitric oxide secretion, and a significant decrease in the production of reactive oxygen species in the hydrolysis of Nanzhu fluid group (P＜0.05). To conclude, hydrolysis of Nanzhu fluid can promote the proliferation and migration of human microvascular endothelial cells in vitro, and has the function of promoting the secretion of nitric oxide and inhibiting reactive oxygen species secretion, implying its positive role in the protection of endothelial cell function.

This study was purposed to investigate the methylation status of id4 gene promoter in patients with chronic myeloid leukemia (CML) and explore the relationship between methylation of the id4 gene and progress of CML. The methylation status of id4 gene in 48 chronic myeloid leukemia patients and 10 healthy individuals was detected by using methylation-specific polymerase chain reaction (MS-PCR). The results showed that id4 gene was unmethylated in bone marrow samples from both healthy individuals and CML patients in chronic phase (CP). The rate of id4 gene methylation in both CML patients in accelerated phase (AP) and blast crisis (BC) was 66%, and was higher than those of CML patients in CP phase. There was significant difference between them (p < 0.05). In one CML patient who received a serial observations, the status of id4 was unmethylated in CP, but it was methylated in AP and BC phase. It is concluded that the id4 gene in CML patients is unmethylated in CP, while it is methylated in AP or BC. The detection of id4 gene methylation status may be useful for monitoring disease advance in CML and may be used as a marker of disease progression in CML.
Adolescent ; Adult ; Aged ; Child ; DNA Methylation ; DNA Primers ; Female ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Male ; Middle Aged ; Promoter Regions, Genetic ; Young Adult