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Objective@#To investigate the effects and mechanism of Holothurian Glycosaminoglycan (hGAG) alone in combination with cisplatin (DDP) on apoptosis of pulmonary adenocarcinoma cell A549.@*Methods@#A549 cells were separately treated with blank, hGAG, DDP and hGAG combined with DDP (hGAG + DDP). The cell morphology in 4 groups was observed using light microscope. CCK8 assay was used to determine the cell viability. Flow cytometry by Hoechst 33258 and AnnexinV-FITC/PI staining was applied to detect cell apoptosis. Western blot was then used to detect the protein expression of Bax, Bcl-2, survivin and caspase-3.@*Results@#After treatment for 24 h, the inhibitory rates of A549 cells in control, hGAG, DDP and hGAG + DDP groups were 0, (19.74±5.39)%, (42.01±2.57)% and (53.89±4.58)%, respectively. Moreover, after treatment for 48 h and 72 h, the inhibitory rates in each group were 0, (23.17±4.78)% and (29.17±4.21 )%, (54.00±7.64)% and (59.35±7.31)%, as well as (77.58±4.26)% and (79.94±4.58)%, respectively. The cell viability was significantly lower in drug treatment groups compared with those in control group at the same time point (P<0.05). Hochest 33258 staining showed that no obvious apoptotic cells were detected in the control group, while apoptotic cells were visible in hGAG, cisplatin and combination groups. Flow cytometry showed that cell apoptotic rates were (2.38±0.59)%, (12.59±4.22)%, (16.36±3.63)% and (44.60±5.45)% in the control, hGAG, DDP and hGAG + DDP groups, respectively. The cell apoptosis was significantly lower in drug treatment groups compared with those in control group at the same time point (P<0.05). Furthermore, western blot results showed that the expression of Bax and caspase-3 protein was increased (P<0.05), whereas Bcl-2 and survivin was decreased (P<0.05) in the hGAG+ DDP group compared with cisplatin alone (P<0.05).@*Conclusions@#HGAG can inhibit the proliferation and promote the apoptosis of human lung adenocarcinoma A549 cells. Meanwhile, it can strengthen the chemosensitivity of A549 cells to DDP via up-regulation of Bax, caspase-3 and down-regulation of Bcl-2 and survivin.
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Objective To investigate the expression of guanylate binding protein 5 (GBP5) and ArfGAP with SH3 domain ankyrin repeat and PH domain 1 (ASAP1) genes in pulmonary tuberculosis patients and tuberculosis latent population. Methods Forty pulmonary tuberculosis patients (pulmonary tuberculosis group), 40 latent tuberculosis infection patients (latent tuberculosis infection group) and 40 cases of healthy control (healthy control group) were selected from August 2016 to May 2017. The gene expression was detected in 4 ml peripheral anticoagulant blood by real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR) and the relative expression of two genes in three groups were compared. Results The GBP5 gene expression in three groups was significantly differentce (F=7.23, P=0.001). The GBP5 gene relative expression in pulmonary tuberculosis group was significantly higher than that in latent infection group :1.58 ± 0.80 vs. 1.09 ± 0.68, there was significant difference (t=2.93, P=0.004). The GBP5 gene relative expression in pulmonary tuberculosis group was significantly higher than that in healthy control group: 1.58 ± 0.80 vs. 1.04 ± 0.61, there was significant difference (t=3.40, P=0.010). The GBP5 gene relative expression in latent infection group and healthy control group had no significant difference (t=0.39, P=0.700). There was no significant difference in ASAP1 expression among three groups (F=0.26, P=0.770). Conclusions The expression of GBP5 in pulmonary tuberculosis patients, latent tuberculosis infection patients and healthy controls is different, and GBP5 could screen latent tuberculosis infection patients which is expected to be a potential screening marker for latent tuberculosis infection.
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Objective To investigate the inhibitory effects of radioactive 125I seeds on the growth and apoptosis of human lung adenocarcinoma cells A 549 in nude mice.Methods Human lung adenocarcinoma A549 cells were cultured in vitro and subcutaneously transplanted in BALA/c nude mice.When the tumor size reached(300 ±50)mm3,40 tumor-bearing mice were divided into 4 groups by the random number table method as 0,0.6,0.8 mCi(1 Ci＝3.7×1010Bq)groups and blank control group,with 10 in each group.The 125I seeds of 0,0.6,and 0.8 mCi were implanted into the transplanted tumors in nude mouse,respectively.The blank control group received no treatment.The weight of nude mice was measured regularly every 4 days.The mice were sacrificed on the 32 days after 125I seeds implication.The transplanted tumors were weighed and the weight gain curve for nude mice was plotted.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of the tumor tissue.Cell apoptosis was detected by TUNEL assay,and the expressions of the P21,Caspase-9,Survivin and Livin proteins were detected by immunohistochemical assay.Results There was no nude mice dead in each group.On the day 28 and 32 after 125I seeds treatment,the body weights of nude mice of 0.6 and 0.8 mCi groups became lighter than those of the blank control group(q＝4.26,9.19,4.11,11.59,P＜0.05),the tumor weights of the 0.6 and 0.8 mCi groups were significantly decreased(q＝5.021,5.692,P＜0.05)with tumor inhibition rates of about 49％and 62％.In the 0.6 and 0.8 mCi groups,a large number of tumor cells degenerated to be necrotic cells.In addition,the apoptotic indexes were(50.00 ±2.58)％and(62.33 ± 4.51)％in the 0.6 and 0.8 mCi groups,respectively,and higher than that of blank control group(27.00 ±4.69)％.The expressions of P21 and Caspase-9 proteins in the 0.6 and 0.8 mCi groups were significantly higher than that in the blank control group(χ2＝11.380,24.310,11.380,20.376,P＜0.05).The expressions of Survivin and Livin proteins in the 0.6 and 0.8 mCi groups was significantly lower than that in the blank control group(χ2＝9.643,23.254,15.429,26.667,P＜0.05).Conclusions Radioactive 125I seeds can inhibit the proliferation of tumor cells and promote the apoptosis of A 549 cells probably by up-regulating the expressions of P21 and Caspase-9 but down-regulating the expressions of Survivin and Livin.
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Objective To investigate the apoptosis and toxicity of oncolytic virus H101 combined with radiation on apoptosis of A549 lung adenocarcinoma cells. Methods A549 lung adenocarcinoma cells in exponential growth phase were divided into four groups: control ( PBS) group, radiation ( IR) group, oncolytic virus (H101) group and radiation combined with oncolytic virus (IR+H101) group. The cells were double dyed with Annexin fluorescein isothiocyanate ( V-FITC/PI ) and then the apoptosis ratio of cells in every group was detected by the flow cytometry. The cytotoxic effect of cells in every group was detected by lactate dehydrogenase ( LDH) release test. The mRNA expression of oncolytic viruses H101 capsid protein Hexon was detected by real-time fluorescence PCR ( RT-PCR) to compare the oncolytic virus replication in each group. Results Cell apoptosis rate in H101 group (55. 37%) was significantly higher than that in PBS group (1.03%) (t =36.51, P <0.05). Cell apoptosis rate in IR + H101 group (93. 06%) was significantly higher than that in H101 group (55. 37%), IR group (12. 67%) and PBS group (1. 03%) (t=13. 51, 24. 14, 38. 99, P<0. 05). LDH releasing percentage in IR group and H101 group at different time after virus transfection was significantly higher than that in PBS group ( t=25. 84,39. 38, 32. 51, 78. 18, P<0. 05;t=31. 40, 2. 68, 23. 43, 60. 98, P<0. 05). LDH releasing percentage in IR+H101 group was significantly higher than that in PBS group (t=80. 71, 119. 74, 109. 80, 123. 94, P<0. 05), IR group (t=28. 80, 54. 34, 72. 34, 61. 91, P<0. 05) and H101 group (t=42. 02, 57. 45, 57. 01, 58. 83, P<0. 05). Compared with H101 group at the same time point, the mRNA expression of Hexon in IR + H101 group at 24, 48 and 72 h was increased by 16. 26, 28. 37 and 39. 58 times, respectively (t=54. 50, 33. 73, 29. 28, P<0. 05). Conclusions The oncolytic virus H101 plays a role of radiosensitization in tumor cells. Radiation also increases the replication of the oncolytic virus H101 and thereby enhances the oncolytic effect of the oncolytic virus H101. Therefore, oncolytic virus H101 combined with radiotherapy has synergistic effect on killing tumor cells.
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Objective To investigate the effect and underlying mechanism of radioactive 125I seed implantation on the angiogenesis of transplanted human lung adenocarcinoma in nude mice.Methods An animal model of transplantd human lung adenocarcinoma was established by subcutaneous implanting A549 cells into nude mice.Twenty four tumor-bearing nude mice were randomly divided into 4 groups with different irradiation doses of blank control (without any treatment) and 0 MBq,22.2 MBq,29.6 MBq and by embedding radioactive 125I seeds with an 18 G implant needle.Tumor volumes were measured every 4 days until all mice were terminated 30 d later and the tumor growth curve was drawn.The microvessel density (MVD) in the tumor tissue was detected by immunohistochemistry S-P assay.The mRNA and protein levels of VEGF and HIF-1α of each group were detected by RT-PCR and Western blot,respectively.Results After embedding of 125I seeds,the tumor volumes of 22.2 MBq group (886 ± 97) and 29.6 MBq group (590 ± 107) were significantly smaller than those of control group (2 297 ± 149) at 54 d after administration (q =14.117,17.075,P ＜ 0.05),but there were no significant differences among 0 MBq group and control group,22.2 MBq and 29.6 MBq groups (P ＞ 0.05).The immunohistochemical CD34-positive staining demonstrated that MVD in 22.2 MBq group (522 ± 119) and 29.6 MBq group (491 ± 121) were decreased significantly compared with control group (922 ± 260) (q =4.826,5.197,P ＜0.05),but there were no significant differences among 0 MBq and control groups,22.2 MBq and 29.6 MBq groups(P ＞0.05).The mRNA expressions of VEGF and HIF-1α in 22.2 MBq group (0.279±0.0659,0.370 ±0.0857) and 29.6 MBq group (0.215 ±0.0620,0.278 ±0.0651) were significantly lower than those in the control group (q VEGFmRNA =18.881,17.211,q HIF-1αmRNA =15.376,14.733,P ＜0.05),but there were no significant differences among 0 MBq and control groups,22.2 MBq and 29.6 MBq groups(P ＞0.05).At the same time,the expression levels of VEGF and HIF-1α protein after 125I seed implantation were also obviously decreased in 22.2 MBq and 29.6 MBq groups (qvEGr =5.848,6.263,q HIF-1α =6.560,7.576,P ＜ 0.05),and no significant difference between 0 MBq and control groups(P ＞ 0.05) and between 22.2 MBq and 29.6 MBq groups (P ＞ 0.05).Conclusions Interstitial implantation with 125I seeds may potently inhibit angiogenesis in human lung adenocarcinoma xenografts of nude mice.
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Objective To investigate the apoptosis and toxicity of oncolytic virus H101 combined with radiation on apoptosis of A549 lung adenocarcinoma cells. Methods A549 lung adenocarcinoma cells in exponential growth phase were divided into four groups: control ( PBS) group, radiation ( IR) group, oncolytic virus (H101) group and radiation combined with oncolytic virus (IR+H101) group. The cells were double dyed with Annexin fluorescein isothiocyanate ( V-FITC/PI ) and then the apoptosis ratio of cells in every group was detected by the flow cytometry. The cytotoxic effect of cells in every group was detected by lactate dehydrogenase ( LDH) release test. The mRNA expression of oncolytic viruses H101 capsid protein Hexon was detected by real-time fluorescence PCR ( RT-PCR) to compare the oncolytic virus replication in each group. Results Cell apoptosis rate in H101 group (55. 37%) was significantly higher than that in PBS group (1.03%) (t =36.51, P <0.05). Cell apoptosis rate in IR + H101 group (93. 06%) was significantly higher than that in H101 group (55. 37%), IR group (12. 67%) and PBS group (1. 03%) (t=13. 51, 24. 14, 38. 99, P<0. 05). LDH releasing percentage in IR group and H101 group at different time after virus transfection was significantly higher than that in PBS group ( t=25. 84,39. 38, 32. 51, 78. 18, P<0. 05;t=31. 40, 2. 68, 23. 43, 60. 98, P<0. 05). LDH releasing percentage in IR+H101 group was significantly higher than that in PBS group (t=80. 71, 119. 74, 109. 80, 123. 94, P<0. 05), IR group (t=28. 80, 54. 34, 72. 34, 61. 91, P<0. 05) and H101 group (t=42. 02, 57. 45, 57. 01, 58. 83, P<0. 05). Compared with H101 group at the same time point, the mRNA expression of Hexon in IR + H101 group at 24, 48 and 72 h was increased by 16. 26, 28. 37 and 39. 58 times, respectively (t=54. 50, 33. 73, 29. 28, P<0. 05). Conclusions The oncolytic virus H101 plays a role of radiosensitization in tumor cells. Radiation also increases the replication of the oncolytic virus H101 and thereby enhances the oncolytic effect of the oncolytic virus H101. Therefore, oncolytic virus H101 combined with radiotherapy has synergistic effect on killing tumor cells.
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Objective To analyze the results of screening for tuberculosis (TB) in health examination participants and study the spontaneous occurrence of TB in the subjects with strong positive reaction to tuberculin skin test (TST) in ten years. Method Totally 12 598 health examination participants without past TB history were selected, of whom 8 896 were college students, 2 496 migrant workers and 1 206 had close contacts with active TB patients. They were screened by TST with strong positive reaction. All of subjects with TST strong positive results received chest X-ray examination and sputum acid-fast bacteria detection. The subjects diagnosed to have TB were given regularly anti-tuberculosis drugs treatment and followed up for ten years. The 429 subjects without TB and no isoniazid preventive treatment were chosen to be followed up for ten years and spontaneous occurrence of TB in first three years and the fourth to tenth years respectively, as well as the recurrence of TB for the patients who received anti-tuberculosis regimen were recorded. Result Thirty-seven cases were diagnosed as TB by TST screening, and the total detection rate was 0.29% (37/12 598). Among them 11 were college students (0.12%, 11/8 896), 12 were migrant workers (0.48%, 12/2 496) and 14 were close contacts (1.16%, 14/1 206) respectively. The detection rates were different among the three groups (χ2=31.40, P=0.000). Among 897 strong positive subjects, the strong positive rate was 7.12%(897/12 598), 316 were college students (3.55%, 316/8 896), 388 migrants workers (15.54%, 388/2 496), and 193 close contacts (16.00%, 193/1 206) respectively. There was significant difference in strong positive rate among the three groups (χ2=583.04, P=0.000), and the strong positive rate of college students was lower compared with that of the migrant workers and the close contacts (χ2=483.51 and 344.11, P<0.01). In ten years, 54 subjects were diagnosed as TB in 429 subjects with strong positive reaction to TST, the spontaneous cumulative incidence rate was 12.58% (54/429). Among them, the cumulative morbidity rate was 9.21%(14/152) in college students, 9.58%(18/188) in migrant workers and 24.72%(22/89) in close contacts respectively. The spontaneous morbidity rate of close contacts was higher than that of college students and migrant workers(χ2=10.63 and 11.21, P<0.001); 75 were lost in 398 participants, the dropout rate was 18.84%(75/398). In first three years of follow-up, 31 were diagnosed TB in 429 participants, the cumulative incidence rate was 7.23% (31/429). Of them 9 were college students (5.92%, 9/152), 10 migrant workers (5.32%, 10/188) and 12 close contacts (13.48%, 12/89) respectively (χ2=6.60, P=0.037). In the fourth to tenth years of follow-up, 23 were diagnosed TB in 398 participants, the cumulative incidence rate was 5.78% (23/398), which was not significantly different compared with the cumulative incidence rate of the first three years (χ2=2.50, P=0.37). Tirty-seven patients received standard anti-tuberculosis drug therapy for one year, no one of them had recurrence at ten-year follow-up. Conclusion The migrant workers and close contacts are the high-risk populations for TB. All of them with strong positive response to TST results are susceptible to TB. So regular physical examination is recommended for them and health management should be strengthened.
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Objective To examine the 5-year incidence of tuberculosis in different populations with strongly positive tuberculin skin test (TST) receiving preventive administration of isoniazide.Methods A total of 12 598 subjects including 8 896 college students,2 496 migrant workers and 1 206 close contacts with active pulmonary were selected from January to December 2003.Subjects with strongly positive TST but without abnormal lung findings were divided into isoniazide group (given isoniazide for 10 months) and control group (not given any drugs).The incidence of tuberculosis in year 2-5 were observed and compared usingx2 test.Results Among 12 598 subjects,897(7.12％) had strongly positive TST,including 316 college students,388 migrant workers and 193 close contacts,and the TST strongly positive rates were 3.55％ (316/8 896),15.54％ (388/2 496) and 16.00％ (193/1 206),respectively.Migrant workers and close contacts had higher TST positive rates than college students (x2 =483.51 and 344.11,P ＜ 0.01).Among 897 TST-positive individuals,37 were diagnosed as tuberculosis,including 11 college students,12 migrant workers and 14 close contacts,and the tuberculosis rates in three populations were 0.12％ (11/8 896),0.48％ (12/2 496) and 1.16％ (14/1 206),respectively.Migrant workers and close contacts also had higher tuberculosis rates than college students (x2 =12.34 and 42.18,P ＜0.01).In the second follow-up year,9 out of 429 subjects in isoniazide group quit the study due to adverse reactions,and in the rest 420 subjects,9 (2.14％) were diagnosed as tuberculosis.The incidence of tuberculosis in three populations were 1.34％ (2/149),1.60％ (3/188) and 4.82％ (4/83),and no significant difference was found (x2 =2.92,P ＞ 0.05).While in the control group,31 out of 429 (7.23％) individuals were diagnosed as tuberculosis,and the incidence was higher than that in isoniazide group (x2 =12.69,P ＜ 0.01).During the next three follow-up years,23 individuals in isoniazide group drop out of the study,and in the rest 388 subjects,8 (2.06％) were diagnosed as tuberculosis.The incidence of tuberculosis in three populations were 1.41％ (2/142),2.35％ (4/170) and 2.63％ (2/76),and no significant difference was found (x2 =3.11,P ＞ 0.05).While in the control group,17 out of 398 (4.27％) subjects were diagnosed as tuberculosis,and the incidence was not of significant difference compared with that in isoniazide group (x2 =2.47,P ＞ 0.05).Conclusion Migrant workers and close contacts are high risk populations of tuberculosis,and preventive administration of isoniazid for 10 months may reduce the incidence of tuberculosis in the following 2 years.
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Objective To study the differential diagnostic value of interferon-γ inducible protein 10 (IP-10),macrophage inflammatory protein-1 α (MIP-1 α) and monocyte chemoattractant-1 (MCP-1) level in the tuberculous,malignant pleural effusion.Methods Enzyme-linked immunosorbent assay was used to detect the level of IP-10,MIP-1 α and MCP-1 in tuberculous pleural fluid (tuberculous pleural fluid group,43 cases) and malignant pleural fluid (malignant pleural fluid group,45 cases).The level of IP-10,MIP-1 α and MCP-1 and the significance were analyzed by ROC curve.Results The level of IP-10,MIP-1 α and MCP-1 were significantly higher in tuberculous pleural fluid group than those in malignant pleural fluid group,and there were significant differences(t =4.931,3.106,2.385 ; P =0.000,0.004,0.041).ROC curve analysis showed that the critical value of IP-10,MIP-1 α and MCP-1 in diagnosis of pleural effusion was respectively 1 589.73,213.50,1 452.63 ng/L.The sensitivity and specificity of IP-10,MIP-1 α and MCP-1 in pleural fluid were 68.8％,81.3％,87.5％ and 87.5％,68.8％,56.3％,respectively.Conclusion The level of IP-10,MIP-1 α and MCP-1 in tuberculous and malignant pleural fluid are significant for the early diagnosis and differential diagnosis.
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ObjectiveTo discuss the clinical diagnosis and treatment of multiple endocrine neoplasia ( MEN ) 2A, and report the mutation of the RET proto-oncogene in a pedigree of three patients with MEN 2A.MethodsBilateral adrenalectomy was performed on two of the three patients with hypertension and bilateral adrenal-conserving adrenal pheochromocytoma resection was performed on the other patient. All three patients were treated by total thyroidectomy and neck lymphadenectomy. Twelve family members were recruited to the study. Peripheral blood was collected and total genomic DNA was prepared for polymerase chain reaction (PCR). PCR products of exon 10 and exon 11 of the RET proto-oncogene were purified and a direct DNA sequence analysis was performed.ResultsThe pathological diagnosis of the specimens was bilateral adrenal pheochromocytoma and medullary thyroid carcinoma in all the three patients. There was no tumor recurrence or distant metastasis after 1.5 - 5 years of follow-up. A missense mutation of TGC (Cys)to CGC (Arg) at codon 634 in exon 11 of the RET proto-oncogene was detected in all three patients. Genetic screening identified two mutation carriers in the other members of this pedigree.ConclusionGenetic mutation screening and surgical intervention may be helpful to the members of high-risk families.
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It is purposed to analyze therapeutic effectiveness at community and designated hospitals for extensive influenza A H1N1 outbreaks at five universities in Qingdao,Shandong,east China during September 1 to 30,2009,including 28 confirmed cases,182 suspected cases and 136 close contacts treated at community health-care facilities,and 56 confirmed cases treated at designated hospitals.There was no significant difference in therapeutic effectiveness between community health-care facilities and designated hospitals (P ＞ 0.05).No new cases occurred seven days after their isolation was lifted for suspected cases and close contacts at community,indicating influenza A H1N1 could be prevented,controlled and cured and its mild cases could be treated at community health-care facilities.
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@#ObjectiveTo explore the effect of early rehabilitation on patients with severe traumatic brain injury.Methods89 patients with severe traumatic brain injury were divided randomly into the treatment group (49 cases) and control group (40 cases). All patients of two groups were treated with routine nursing, dehydrated drug, brain protective therapy and alimental therapy, and measures of preventing complications. While, the patients of the treatment group were added with motor and cognitive rehabilitation, mainly physical therapy, combined with occupational therapy, psychotherapy and speech therapy. All patients of two groups were evaluated with scores of activities of daily living (ADL), Disability Rating Scale (DRS), Fugl-Mayer Assessment (FMA), Mini-mental status examination (MMSE), three class balance scale before and after treatment.ResultsThe scores of ADL, DRS, FMA and balance function of the patients in the treatment group were significantly different from that of the control group after treatment ( P<0.05).The MMSE scores of two groups had no significantly difference after treatment ( P>0.05).ConclusionEarly rehabilitation can improve the brain function of the patients with severe traumatic brain injury, patients' living quality and ADL.
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To propose the hypothesis of the requirement for optimum ratio of various active cons tituents of Chinese materia medica with desired effects, which suggests that both single drug and compound drug prescriptions have their own target effects corresponding to their main indications and their active constituents must be in optimum ratio This hypothesis was initially tested in an ischemic animal model The neuroprotection of Yushen Oral Liquor (YSOL, an oral liquor made of Chinese materia medica) on neuronal injury in CA1 region of rat hippocampus following transient forebrain ischemia was used as an index to evaluate the drug's effect By combination of HPLC and pharmacological method, the relationship of HPLC and pharmacodynamical results of each sample made of YSOL was studied to observe the chromatogram peaks representing the drug's neuro protective effects There are three peaks named as R1, R2, and R3 for YSOL, which closely correlate to the drug's neuronal protective effects and the relative weight but not the absolute quantity of those peaks plays a critical role in producing neuronal protective effects of YSOL on ischemia induced neuronal damage The results provide experimental data supporting our proposed hypothesis