ObjectiveTo study the efficacy of using multileaf collimators with different position and different degree in the treatment of nasopharyngeal carcinoma (NPC) using intensity-modulated radiotherapy techniques.Methods Ten patients withNPC were administered andanalyzed.Thepenumbra characteristics, dose of target, and radiation conformal indexes (CI) of mode T1 and mode T2 were measured and compared using dose volume histogram generated by Varian Eclipse three-dimensional planning computer system. Mode T1 :The angles of seven coplanar beams were 0°, 52°, 106°, 160°, 212°, 258°and 308°,respectively. There were no restriction on the position and degree of multileaf collimators. Parameters were set and optimized. Mode T2 :The beam angles and the parameters were as same as mode T1. According to the actual situations, the position and the degree of the multileaf collimators were changed. Then thedose optimization was performed. Results Target dose coverage in both mode T1 and T2 could be clinically accepted, and the CI were 0. 82 and 0. 83(t = -0. 25, P =0. 815). The maximum dose reductions in the lens, eyes, optic nerves and corneas were 28. 7% (t = 4. 80, P = 0. 000), 2. 7% (t = 2. 99, P = 0. 021),1.4%(t= 1.05,P=0.032), and 30.5% (t=2.99,P=0. 020), respectively. However, the mean dose and V35 of the parotid were increased by 0. 6% (t = - 2. 82, P = 0. 043) and 9.9% (t = - 2. 05, P =0. 038). ConclusionsOpimization of multileaf collimators can reduce the scattering and leaking rays. Compared with mode T1 ,controlling the position and degree of multileaf collimators could reduce the radiation dose to the eyes and optic-nerves, especially to the lens.

Objective To establish a universal stem loop primer (USLP) based real-time PCR method to scan mature miR profile and quantify it's expression.Methods The common universal stem-loop primer pairs were re-designed; 8 random nucleotides were introduced at 3 ' end for reverse transcription of the mature miR,establishing a miR scanning and quantifying system based on SYBR Green Ⅰ PCR (improved USLP method).10-fold gradient diluted standard miRNA-155 cDNA ( 1 ～ 109 copies/μ1) were utilized to evaluate the sensitivity of this method.The specificity was verified by melting curve assay; the precision was assessed by intra-assay coefficient of variation (ICV) of threshold cycle (Ct value) through 20 repeated detections of the standard miR-155 cDNA (2 × 105,2 × 106,2 × 107 copies/μl) ; cost of the primers and time were evaluated,compared with that of the conventional USLP method.Peripheral blood samples were cultured with phytohaemagglutinin (PHA) for0 h,16 h,24 h,48 h and 72 h,and 87 candidate miR that may be associated with human immunity from PubMed data were scanned and quantified from the cultured T cells.Results The sensitivity of the improved USLP method was 103 copies/μl of standard miR-155 cDNA.Melting curve assay showed a single melting peak at 80 ℃,suggesting the excellent PCR specificity of miR-155.Precision of our method quantifying miR-155 was acceptable (ICV ＜ 2.5％ ).Compared with the traditional stem loop primers,our method saved 75％ cost of primers ( 1 917 bp vs 7 851 bp) and 60％ test time of reverse transcription (85 min vs 205 min).By our method,85 of the 87 miR expression in T cells had no significant difference after the PHA stimulation; the expression of miR-150 (72 h) decreased by 10 times and that of miR-155 (48 h) increased 8 times after culture with PHA (Z =-2.032,P =0.042;Z =- 2.023,P =0.043,respectively ).Conclusions The improved USLP method is fast,precise,sensitive,and cost-effective.It could be used for miR profile scanning and quantifying in T cells.

OBJECTIVE: To observe the effects of melittin on the expressions of mitochondria membrane protein 7A6, cell apoptosis-related gene products Fas and Fas ligand (FasL) in hepatocarcinoma cells in vitro and to study the mechanisms of melittin in inducing apoptosis of hepatocarcinoma cells. METHODS: BEL-7402 cell line was treated with melittin in vitro. The expressions of mitochondria membrane protein 7A6, cell apoptosis-related gene products Fas and FasL were detected by flow cytometry. Fas and FasL mRNAs were analyzed by reverse transcription polymerase chain reaction (RT-PCR) assay. RESULTS: The expression rates of mitochondria membrane protein 7A6 of BEL-7402 hepatocarcinoma cells in 8, 16, 32 microg/ml melittin-treated and control groups were 4.89%, 17.74%, 11.45% and 1.02%, respectively. The expression of Fas protein was up-regulated by melittin, while FasL expression did not change. RT-PCR results showed that Fas mRNA expression was up-regulated by 32 microg/ml melittin and FasL mRNA expression was not observed. CONCLUSION: The effects of melittin in inducing hepatocarcinoma cell apoptosis may be related with up-regulating the expressions of mitochondria membrane protein 7A6 and Fas protein.

ObjectiveTo study the role of alpha smooth muscle actin (α-SMA) positive cells (pericytes)in early myocardial ischemia.MethodsThirty healthy female Wistar rats were randomly divided by body weight into normal control group and subcutaneous multi-point injection of isoprenaline(Isp) group.Five rats were anesthetized after 0,2,4,6,12,24 and 48 h in each group.Blood was taken in eyeballs and,serum was separated,heart was taken and fixed in 4％ paraformaldehyde solution.The myocardial enzymes [serum apartate aminotransferase ( AST ),lactate dehydrogenase (LDH),creatine kinase (CK),creatine kinase isoenzymes (CK-MB)] were determined by biochemical automatic analyzer,the expression of α-SMA and vimentin in myocardial tissue was detected by immunohistochemical staining and changes of pericytes in early myocardial ischemia was observed by morphometric analysis.ResultsThe level of myocardial enzymes AST[(160.25 ± 3.86),(172.60 ± 8.82),(192.20 ± 25.35)U/L],and LDH[(1466.25 ± 643.38),(1645.20 ± 326.83),(1701.60 ± 640.06)U/L],12,24 and 48 h after subcutaneous injection of Isp,were higher than that[(129.18 ± 19.65),(849.45 ± 248.54)U/L,all P ＜ 0.05] of the control group.The level of CK[(1097.60 ± 301.57),(1247.20 ± 243.84),(1263.75 ± 271.22),(1448.00 ± 647.00),(1268.40 ± 479.81)U/L],and CK-MB[(217.12 ± 35.89),(229.08 ± 97.11),(251.70 ± 46.82),(318.80 ±77.76),(249.04 ±:98.54)U/L],4,6,12,24 and 48 h after subcutaneous injection of Isp,were higher than that [(713.45 ± 146.30),(147.05 ± 25.75)U/L,all P ＜ 0.05] of the control group.The number of α-SMA positive cells(61.00 ± 17.25),4 h after Isp injection,was significantly increased compared with that(28.20 ± 5.81,18.20 ± 2.17) of 24 and 48 h after Isp injection.The number of α-SMA positive cells in 48 h group was less than that(50.00 ± 3.61,P ＜ 0.05) of 6 h group.The number of vimentin positive cells in 6,12,24,48 h groups (4.17 ± 2.49,5.24 ± 2.84,8.37 ± 2.18,7.73 ± 2.49) were higher than that(1.88 ± 1.85,2.21 ± 1.54) of the control group and 4 h group(all P ＜ 0.05).Compared with 24 and 48 h groups,the level of vimentin protein was increased in 6 and 12 h groups(P ＜ 0.05).ConclusionThe number of pericytes in early myocardial ischemia is higher than that of other mesenthymal cells,and pericytes may be the main effector cells in the generation and development of myocardial fibrosis.

Objective To compare the efficacy and complications between an auto negative pressure generation biopsy device and Bard biopsy device in renal biopsy in the elderly.Methods A total of 282 patients in our department received renal biopsies with auto negative pressure generation biopsy device (n=159) versus Bard device (n=123).The quality of tissue biopsy specimen and postoperative complications were analyzed retrospectively.Results There were no significant differences in the success rate,incidences of perirenal hematoma and gross hematuria between the two groups (96.9％ vs.95.1％,29.6％ vs.30.1％,1.9％ vs.1.6％,P＞0.05).While the average number of glomeruli,the average length and width of kidney tissue specimen were much more,longer or wider by the auto negative pressure generation renal biopsy device than by the Bard device [(17.9± 11.5) vs.(12.6±9.9),(11.5±5.0)mm vs.(7.8±3.0) mm,(1.0±0.2) mm vs.(0.8±0.4) mm,respectively,all P＜0.05].Conclusions There are no significant differences in the success rate and postoperative complications between auto-negative pressure generation renal biopsy device and Bard device.The auto negative pressure generation renal biopsy device has the advantage in obtaining more renal tissue,with the same effectiveness and safety as the Bard device.

AIM:To investigate the effect of cyclopamine on Hedgehog (HH) signaling, phenotypic transfor-mation and matrix accumulation induced by aristolochic acid (AA) in renal tubular epithelial cell NRK-52E.METHODS:NRK-52E cells were randomly divided into control group (treated with solvent only), AA group (treated with AA at con-centrations of 1, 5, 10 mg/L) and cyclopamine group (treated with AA at concentration of 10 mg/L plus cyclopamine at concentrations of 1, 5, 10μmol/L).After cultured for 24 h, the mRNA expression of Ptch1, Smo,α-SMA, E-cadherin, ZO-1, BMP-7, type I collagen and type III collagen was quantified by real-time PCR.The protein levels of Shh and TGF-β1 were detected by ELISA .Immunofluorescence staining was used to evaluate the expression of Ptch 1, Smo,α-SMA, E-cadherin and type III collagen in the NRK-52E cells.RESULTS: AA increased the expression of TGF-β1, α-SMA and type III collagen, decreased the expression of E-cadherin and ZO-1 protein, and down-regulated the expression of Ptch1, Shh and Smo mRNA in the NRK-52E cells, indicating that AA activated HH signaling , and phenotypic transformation and matrix accumulation occurred in AA-treated NRK-52E cells.Treatment with cyclopamine inhibited HH signaling by decrea-sing Smo expression and increasing Ptch 1 expression.Moreover, cyclopamine also down-regulated the expression of TGF-β1,α-SMA, type I collagen and III collagen , and up-regulated the expression of BMP-7, ZO-1 and E-cadherin.CON-CLUSION:AA induces phenotypic transformation and matrix accumulation in renal tubular epithelial cells , which can be inhibited by cyclopamine treatment .The possible mechanism is that cyclopamine suppresses the activation of HH signaling , resulting in the reduction of epithelial-to-mesenchymal transition and matrix deposition .

Objective To compare the differences between two statistical methods for evaluating non-sensitivity of pathogenic bacteria to antimicrobial agents,and explore effect of non-consideration of clinical background on evalua-ting extent of bacterial resistance.Methods Data of Staphylococcus aureus and Acinetobacter spp .in a hospital in the first half year of 2008,2010 and 2013 were collected and conducted statistical analysis with two methods (me-thod 1 :based on all clinically isolated bacteria;method 2 :based on infection-related non-repetitive bacteria),two methods for evaluating bacterial non-sensitive rates to antimicrobial agents were compared.Results The non-sensi-tive rates of Acinetobacter spp .to various antimicrobial agents :statistical results by using method 1 were generally higher than those using method 2,absolute difference between two statistical methods was 10.46%-33.77%;the non-sensitive rates of Staphylococcus aureus to various antimicrobial agents :except compound sulfamethoxazole in 2010 and 2013(difference were 6.17% and 10.21 % respectively),penicillin G (difference was 3.86%),erythromy-cin (difference was 2.71 %),and azithromycin in 2013 (difference was 2.43%),statistical results by using method 1 were generally higher than those using method 2,absolute difference between two statistical methods was 0-18.04%.Conclusion There are deviation in the non-sensitive rates of bacterial strains to antimicrobial agents by using two different statistical methods,deviation is larger in Acinetobacter spp ..The resistance level might be incorrectly higher when evaluating the resistance status without considering clinical background of bacteria.

Objective To assess the value of diffusion weighted imaging (DWI) and apparent diffusion coefficient (ADC) in differential diagnosis of neuroendocrine carcinoma of the uterine cervix (NECUC) from other tumors.Methods A total of 12 NECUCs,39 cervical squamous cell carcinomas (SCC) and 21 cervical adenocarcinomas (CA) confirmed by pathology were analyzed retrospectively.All the patients underwent conventional MRI and DWI.The ADC values were measured and compared among NECUC,SCC and CA.Diagnostic performance of ADC was compared using receiver operating characteristic curves (ROC).Results The mean ADC values of NECUC,SCC and CA were (0.66 ± 0.11) ×10-3 mm2/s,(0.86 ± 0.11) × 10-3 mm2/s and (1.04 ± 0.17) × 10-3 mm2 / s,with statistical differences between any two groups (P ＜0.001).The optimal cutoff values of ADC for differentiating NECUC and SCC was 0.681 ×10-3mm2/s with a sensitivity of 94.9％,specificity of 75.0％ and accuracy of 90.2％.The optimal cutoff values of ADC for differentiating NECUC and CA was 0.824× 10-3mm2/s with a sensitivity of 95.2％,specificity of 91.7％ and accuracy of 98.9％.Conclusion The differences of the mean ADC value are helpful for the differential diagnosis of NECUC,SCC and CA.

Objective To investigate the expression of monocyte-macrophage-related factors and interstitial fibrosis in kidney tissues of rats with ureter obstruction and recanalization .Methods Forty-eight male Spragur-Dawley rats were divided randomly into the obstructive group:sham (n=6), unilateral ureteral obstruction(UUO)3 days (n=6), UUO 7 days (n=6), and UUO 14 days (n=6) and recanalization group:bilateral ureteral obstruction(RBUO)0 day (n=6), 3 days after RBUO (n=6), 7 days after RBUO (n=6), and 14 days after RBUO (n=6).The kidneys were excised on day 3, 7, and 14, and the deposition of collagen fibers in kidney was detected with HE and Masson staining . Immunohistochemical analysis was performed to evaluate the protein expressions of monocyte chemoattractant protein -1 (MCP-1), macrophage colony-stimulating factor (M-CSF) and activated-macrophage marker CD68.Real-time PCR was used to detect the mRNA expressions of MCP-1 and M-CSF.TGF-β1 levels were determined by ELISA .Results Fibrosis observed with HE and Masson staining was obviously increased in kidney tissue of UUO rats , and aggravated as time prolonged, but alleviated in rats with recanalization .TGF-β1 levels were increased obviously in the UUO group , but decreased in rats with recanalization compared with those in BUO rats .In UUO rats, mRNA and protein expression levels of MCP-1 and M-CSF were increased .MCP-1 and M-CSF expression was gradually decreased in rats with recanalization compared with those in BUO rats .The dynamic change in expression of MCP-1 and M-CSF in both UUO rats and recanalization rats was consistent with the change in expression of CD 68. Conclusion Dynamic change in expression of MCP-1 and M-CSF in kidney tissues reflects change of activated and accumulated monocyte -macrophages , which may be one of the major mechanisms contributing to fibrosis induced by ureter obstruction .Renal fibrosis is alleviated by down-regulated expression of monocyte-macrophages factors with recanalization operation .

Objective To analyse the diagnosis and treatment of testicular torsion. Methods The clinical data of 66 cases of testicular torsion were retrospectively analysed. Results Among the 66 cases,32(48.5%) paid the first medical visit within 10 h,and 24(36.4%)were confirmed diagnosis at the first visit.False negative results occurred with color Doppler flow imaging(CDFI),and 8 testicles were damaged due to the false negative diagnosis.Thirty-three patients without prophylactic contralateral orchidopexy were followed up for 6 months to 20 years,and one experienced recurrent torsion. Conclusion The testicular torsion must be considered when a sudden acute scrotum pain is occurred.Testicular damage is closely related to the torsion time,and delayed medical intervention contributes to the testicular damage.Highly suspected cases should be performed surgical exploration timely due to the false negative results with CDFI.Prophylactic contralateral orchidopexy is recommended.