To investigate the effects of anti-sense peptide nucleic acids (PNAs) targeting Ki-67 gene on modulation of the proliferation and apoptosis of human renal carcinoma cell lines, human renal carcinoma cell line 786-0 cells were treated with anti-sense PNAs at different concentrations (1.0 micromol/L, 2.0 micromol/L, 10.0 micromol/L). The Ki-67 expression of 786-0 cells was detected by immunohistochemical technique and Western blot method respectively. The proliferation of 786-0 cells was studied by cell growth curves and 3H-thymidine incorporation. The apoptosis of 786-0 cells was detected by TUNEL assay. The control groups were treated with anti-sense oligonucleotide (ASODNs) targeting Ki-67 gene. Our results showed that the Ki-67 expression of 786-0 cells treated with anti-sense PNAs (16.9+/-0.7) was significantly inhibited as compared with that of the control groups (28.6+/-0.4) (P<0.01). The Ki-67 protein rate of 786-0 cells treated with anti-sense PNAs (42.1 +/-2.2) was significantly reduced when compared with that of the control groups (83.6+/- 1.4) (P<0.01). Proliferation of 786-0 cells treated with anti-sense PNAs (20.7+/- 1.5) was significantly inhibited as compared with that of the control groups (58.6+/- 1.4) (P<0.01). The apoptosis rate of 786-0 cells treated with anti-sense PNAs (28.7+/- 2.3) was significantly increased higher compared with that of the control groups (13.8 +/- 1.0) (P<0.01). From these finds we are led to conclude that anti-sense PNAs targeting Ki-67 gene have stronger effects on the inhibition of the proliferation and induction of apoptosis of human renal carcinoma cells than ASODNs targeting Ki-67 gene. The strategies using anti-sense PNAs targeting Ki-67 gene may be a promising approach for the treatment of renal cell carcinoma.

Objective To explore the effects of Tongfu Huayu Daotan Decoction (通化阏导痰汤)on the serum concentration of S-100β protein, neuron-specific enolase(NSE) and Prognosis in patients with severe craniocerebral injury.Methods Sixty patients with severe craniocerebral injury were randomly divided into a a'eatment group and a control group.The treated group was treated with Tongfu Huayu Daotan Decoction plus conventional treatments including dehydration,antibiotics, organ functional support, nerve nutrition, prevention of complication, etc.; the control group was treated with conventional treatments alone. The concentration of serum S-100β protein and neuron-specific enolase(NSE) in plasma at admission and at24, 36, 72huors, and 5, 7 days after treatment were determined respectively; the Glasgow outcome scale (GOS)and neurological deficits scoring at 2weeks and 4weeks after hospitalization were compared to observe the efficacy of the patients. Results The concentration of serum S-100β protein and neuron-specific enolase (NSE) in plasma at previous treatment and at 24, 36 hours after the treatment had no statistical difference in the two groups(P＞0.05 ), The concentration of serum S-100β protein and neuron-specific enolase (NSE) in plasma at 72huors, 5 and 7 days after the treatment in the Tongfu Huayu Daotan Decoction group were lower than those in the control group, the differences being significant (P＜0.01). The Glasgow outcome scale (GOS) and neurological deficits scoring at 2weeks and 4weeks after the treatment in the Tongfu Huayu Daotan Decoction group were significantly nigher than those in the control group, the differences being significant (P＜0.01) .Conclusion Tongfu Huayu Daotan Decoction can alleviate the plasma concentrations of S-100β protein and neuron-specific enolase (NSE) in patients with severe craniocerebral injury and markedly improve the clinical therapeutic effects. Combined Tongfu Huayu Daotan Decoction and western medicine can significantly reduce mortality and improve the Glasgow outcome scale (GOS), neurological deficits scoring and therapeutic effect.

Objective To investigate the inhibition effect of Ki67 AS-ODN on pr ostate carcinoma PC-3 cells,and its possible synergism existing in combination of Ki67 AS-ODN and paclitaxel.Methods Ki67 AS-ODN were transfected into PC-3 cells by lipofectamine. Cell proliferation was measured by CCK8 method,and Ki6 7 mRNA expression was detected by RT-PCR. The synergetic effect of Ki67 AS-ODN combined with paclitaxel was evaluated by Zhengjun Jin Q method. Results AS-OD N at the concentration of 31.25 nmol /L can significantly inhibit PC-3 cells pr oliferation(P

Objective:To investigate the anti-tumor immune responses against renal cell carcinoma induced by dendritic cells(DCs)transfected with survivin gene mediated by recombinant adenovirus.Methods:The DCs derived from human peripheral blood were infected with recombined adenovirus vector carrying the survivin gene.The expression of survivin protein in the infected DCs was examined by Western blotting assay;the surface expression of CD83,MHCⅡ,CD80 and CD86 by flow cytometry(FCM),Interleukin-12(IL-12)in the supernatants of DCs and IFN-? released by the cytotoxic T lymphocytes(CTLs)by ELISA,the ability of DCs in proliferating allo-lymphocytes by mixed lymphocyte reaction(MLR),and specific killing activity of CTLs by MTT assay.Results:The DCs presented mature DCs phenotype after transfection with Ad-svv.The expression of survivin protein in transfected DCs was confirmed by Western blotting analysis.The IL-12 level in the supernatant of DCs transfected with Ad-svv was significantly higher than that transfected with empty vector(Ad-CMV,P

Objective To explore the significance of combined detection of D-Dimer,cardic troponin I(cTnI)and N-terminal pro-brain natriuretic peptide(NT-ProBNP)in acute coronary syndrome and the correlation between them.Methods 143 patients with acute coronary syndrome were selected as the observation group,and 40 CAG negative people as the control group.The difference between the two groups was compared and the correlation was analyzed.According to different diagnostics,patients in the observa-tion group were separated into 3 groups,unstable angina pectoris,non ST segment elevation myocardial infarction and ST segment elevation myocardial infarction.The correlation of D-Dimer,cTnI and NT-ProBNP with the severity of coronary artery disease was analyzed.Results The levels of D-Dimer,cTnI and NT-ProBNP in the observation group were higher than those in the control group,and the difference was statistically significant(P < 0.05 ).The severity of coronary artery disease had positive correlation with the test results (P <0.05).Conclusion The combined detection of D-Dimer,cTnI and NT-ProBNP could help to diagnose the acute coronary syndrome and the test result has a positive correlation with the severity of acute coronary syndrome.

BACKGROUND: Percutaneous vertebroplasty (PVP) is usually used for osteoporotic thoracolumbar fractures,which has various advantages such as easy to operate, short operation time, less trauma, rapid recovery,analgesic effect and so on. But its application is restricted due to nerve compression symptoms and pulmonary embolism caused by bone cement leakage. Thereafter, how to reduce the leakage of bone cement is an issue of concern.OBJECTIVE: To investigate the relationship between the lumbar quantitative computed tomography (QCT) values and contrast agent dispersion in osteoporotic thoracolumbar fractures. METHODS: Sixty cases of osteoporotic thoracolumbar fractures undergoing PVP were enrolled, and received QCT examination before surgery, and contrast agent was injected intraoperatively. X-ray examination was conducted to detect the bone mineral density, contrast agent dispersion and leakage of bone cement, and the relationship between the lumbar QCT values and contrast agent dispersion as well as leakage of bone cement.RESULTS AND CONCLUSION: (1) There were 110 vertebral fractures, and 74 vertebrae with contrast agent diffusing more than vertebral midline, accounting for 67.3%. There was significant difference in the contrast agent dispersion among groups (P < 0.05). (2) The bone cement leakage showed no significant difference among groups after injected with bone cement by unilateral or bilateral approach (P > 0.05). (3) These results suggest that contrast agent dispersion in osteoporotic thoracolumbar fractures has a certain relationship with the lumbar QCT values, and lumbar QCT values with more contrast agent dispersion, but the lumbar QCT values have no correlation with bone cement leakage. Therefore, choosing a appropriate approach based on the QCT values and contrast agent dispersion can reduce leakage and improve the safety of PVP.

Objective To express adenosine deaminase protein by molecular cloning technique.MethodsTotal RNA was extracted from human leukocytes and the cDNA was obtained by reverse transcription.Whereupon the cDNA was used as a template to amplify adenosine deaminase gene by polymerase chain reaction (PCR) and then integrated it into three prokaryotic plasmids pET-28 b, pET-32 a (+) and pHSIE.The plasmid with the correct sequencing was transformed into E.coli BL21 (DE3) by CaCl2 method for the protein expression.The expression activity of these fusion proteins were detected by Western-blot and SDS-PAGE, with the optimized expression conditions.Results Complete fusion of target gene and three prokaryotic plasmids was observed through sequencing.The expressed and accurate ADA protein was identified by Westernblot and SDS-PAGE.The optimal expression conditions were observed:the protein expression would be induced with 0.4 mmol/L IPTG and incubated at 16℃for 24 hours.Conclusion The prokaryotic vectors of adenosine deaminase (BL21+pET-28 b+ADA, BL21+pET-32 a+ADA, BL21+pHSIE+ADA) were successfully constructed and efficiently expressed.

OBJECTIVETo establish of blood beta hydroxybutyrate (βOHB) threshold for diagnosing type 2 diabetes ketoacidosis (DKA) and explore the relationship between βOHB levels and the severity of DKA.

METHODSCorrelation analysis was performed between serum βOHB and [HCO(3)] in type 2 diabetic patients admitted in the emergency department in the past year. Regression equation was used to calculate the concentration of βOHB corresponding to a [HCO(3)] level of 18.0, 15, and 10.0 mmol/L, and βOHB concentration corresponding to a [HCO(3)] level of 18.0 mmol/l was used as the DKA diagnostic threshold.

RESULTSThe serum βOHB level and [HCO3] concentration showed a good correlation (R²=0.7023, P<0.001). βOHB concentrations that corresponded to a [HCO(3)] level of 18.0, 15, and 10.0 mmol/L were 3.0, 4.70, and 7.5 mmol/L, respectively, in accordance with the severity of DKA. Combined with the blood glucose concentration ≥ 13.9 mmol/L, a blood βOHB≥3.0 mmol/L showed a sensitivity of 99%, specificity of 86%, and total effectiveness of 92.81% for diagnosing DKA.

CONCLUSIONA serum βOHB level above 3.0 mmol/L can be used as the diagnostic threshold of DKA. βOHB can serve as an index for assessing the severity of DKA.

3-Hydroxybutyric Acid ; blood ; Diabetes Mellitus, Type 2 ; blood ; Diabetic Ketoacidosis ; blood ; diagnosis ; Humans ; Sensitivity and Specificity ; Severity of Illness Index

Objective Clinical study on the treatment of bilateral lumbar spinal stenosis with percutane-ous fixation combined with unilateral open-ended spinal canal decompression. Methods 126 patients with bilater-al lumbar spinal stenosis admitted to our hospital were randomly divided into two groups.The observation group was treated by percutaneous nail combined with unilateral laminar fenestration,and the control group was treated by open reduction combined with bilateral hemi laminectomy and spinal canal decompression.The two groups of pa-tients with general surgical complications after treatment,index,lumbago and leg pain VAS score and ODI score were compared.Results The operation time of the observation group,the amount of bleeding,the time of hospital-ization and the cost of hospitalization were less than those of the control group.There were no complications such as incision infection after operation in the two groups.The two groups were statistically significant postoperative pain and leg pain VAS score and ODI score compared with preoperative difference.The two groups had statistical signifi-cance between low back and leg pain VAS score and ODI score after 6 and 12 months and last follow-up phase dif-ference.But the two groups after 3 months of lumbago and leg pain VAS score and ODI score had no significant dif-ference.Conclusions Percutaneous minimally invasive nail combined with unilateral laminar fenestration and de-compression for bilateral lumbar spinal stenosis has the advantages of less trauma,less bleeding,shorter hospitaliza-tion time and quicker recovery.It is worthy of clinical promotion.

Objective To determine the optimal electron beam energy at different field size through a Monte Carlo-based simulation of the therapy head of Varian X 6 MV linear accelerator so as to study the influence of radial intensity on depth dose.Methods Firstly,keeping the radial intensity unchanged for the field of interest while changing electron beam energy,compassion was carried out of calculated percentage depth doses between measured values.Thus,the optimal energy was identified for this field size.Then,the obtained energy was set the optimal value to study the radial intensity influence on the depth doses.Results The optimal electron energy for 4 cm ×4 cm,10 cm × 10 cm,20 cm × 20 cm and 30 cm × 30 cm field sizes was 5.9,6.0,6.3 and 6.4 MeV respectively.Changes in radial intensities resulted in negligible changes in percentage depth doses for4 cm ×4-cm and 10 cm × 10 cm fields,but led to observable discrepancy for 20 cm × 20 cm and 30 cm × 30 cm fields.Conclusions The optimal electron energies for different field sizes are slightly different.Change in radial intensity distribution has significant influence on the depth dose for large field.To improve simulation accuracy,the field size needs to be taken into consideration in determining the electron beam energy and radial intensity distribution.