Background and purpose:It has been confirmed that homozygous deletion of p16/p15 gene and its co-deletion of p16/p15 genes were related to the occurrence, progress and prognosis of epithelial ovarian cancer. However, the mono-deletion and co-deletion of the genes has been detected with tissue but not in serum DNA of the epithelial ovarian cancer. In this article, we studied the relationship between homozygous deletion of p16/p15 gene and its co-deletion of p16/p15 genes in serum DNA of the epithelial ovarian cancer.Methods:Primers were used to amplify exon 2 of p16 and exon 2 of p15 gene by polymerase chain reaction. Homozygous deletions of the p16, p15 and co-deletion of p16/p15 genes were studied in either serum DNA of 165 patients with epithelial ovarian cancer, their counterpart lymphocytes DNA, serum DNA of 25 benign ovarian cyst or of 15 health donors.Results:The homozygous deletion rates of either p15 or p16 gene were 27.9%(46/165)and 27.3%(45/165)serum DNA in the patients with epithelial ovarian cancer respectively, while the co-deletion rate of p16/p15 genes was 24.2% (40/165). However, the deletions of p15/p16 genes and its co-deletion were not found in serum DNA of the counterpart lymphocytes,25 benign ovarian cyst and 15 health donors (The P values were 0.000、0.000 and 0.000 respectively). The deletions of either p15 or p16 gene for the patients with stage Ⅰ~Ⅱ were 14.3%(5/35) and 11.4%(4/35), 33.3%(25/75) and 32.0%(24/75) for the patients with stage Ⅲ, 29.1%(16/55) and 30.9% (17/55) for stage Ⅳ, respectively. Although there was no significant differences among the groups, the deletion of p15 and p16 genes in the patients with advanced stage were higher than that with early stage. The deletion was not found to be associated with histopathology of epithelial ovarian cancer.Conclusions:Homozygous deletions of the p16, p15 genes and its co-deletion of p15/p16 genes were commonly found in the serum DNA of epithelial ovarian cancer and might be associated with clinical stage of the disease. It was suggested that detection with serum DNA may be used as a micro-invasive approach and the deletion of genes might served as biological markers for the development and prognosis of the patients with epithelial ovarian cancer.

Objective Investigate the frequency of loss of heterozygosity (LOH) at chromosome arm the short arm chromosome 3p14,25 in the serum DNA from ovarian cancer and its clinical application Methods Thirty eight ovarian cancer serum samples with 18 corresponding tumor tissues and 8 benign ovarian tumors were obtained,and DNA samples extracted from serum and tissue were examined for 3p14,25 LOH by using of polymerase chain reaction with four polymorphic microsatellite markers (D3S1029, D3S1228, D3S1300, D3S1481) Results Matched serum and tissue DNAs from 18 ovarian cancer patients showed significant concordance of 3p14,25 LOH ( P

We prepared gold nanostar (GNS) through seed growth method firstly,then formation of COX-2 siRNA(siCOX-2) and GNS composite modified with polyethylene glyco (PEG),2-amino-2-deoxy-D-glucos (DG) and 9-D-arginin (9R) was prepared.Mterwords,paclitaxel temperature sensitive liposome (PTX-TSL) was prepared by film dispersion method.Finally,siCOX-2 delivery systerm (PTX-TSL-(siCOX-2(9R/DG-GNS)))was obtained by hydrosulfuryl ligand reaction between siCOX-2 (9R/DG-GNS) and PTX-TSL The successful build of siCOX-2 (9R/DG-GNS) was vetified by nuclear magnetic resonance (NMR),sodium dodecyl sulfate polyacryl amide gel electrophoresis (SDS-PAGE),and ultraviolet spectrophotometry and agarose gel electrophoresis method.Particle size of PTX-TSL-(siCOX-2(9R/DG-GNS)) was (292 ± 14) nm and Zeta potential was about -(2.59 ± 0.12) mV,which were measured by Zetasizer Nano ZS90.The morphology of PTX-TSL-(siCOX-2 (9R/DG-GNS)) measured by transmissionelectronmicroscopy showed homogeneous star structure with phospholipid bilayer on the surface,and it showed good thermal conversion efficiency under radiation of 808 nm laser.Differential scanning calorimetry test showed that PTX-TSL phase transition temperature is about 42.6 ℃.The drug loading content(using dialysis method) and encapsulation efficiency of PTX-TSL were about 7.5％ and 95.4％,at the same time,the release process experiment of PTX-TSL showed that it had a good temperature sensitive release performance.It is hopeful that this siCOX-2 system can be used for reducing drug resistance of PTX and improving the treatment effect of PTX through the synergistic antitumor drug resistance effect of siCOX-2.

10.3969/j.issn.1007-3969.2013.05.009

Objective To investigate the preventive effect of nano-powder of Wugong-sanqi (NW),the rhizome of Anemone flaccid,on postoperative intestinal adhesion in rats.Methods Fifty SD rats were subjected to operation with Ellis' method for establishing intestinal adhesion models,then randomly divided into 5 groups (n=10),namely model,positive (Dexamethasone,i.m.10 mg/kg),NW high,medium and low dose group (p.o.450,225 and 112 mg/kg,respectively).Another ten normal rats were selected as normal control group.After administration 3 days pre-operation and 7 days post-operation,all of rats were killed,the intestinal adhesion was graded and the tissues were observed by optical microscope.Results NW evidently reduced the severity of postoperative adhesion (P＜0.05 or P＜0.01),compared with model group.The histopathologic changes such as proliferation of fibroblast cells and capillary,interstitial granulomas and inflammatory cells infiltration in intestinal tissues were also improved significantly in NW groups.Conclusion NW could inhibit the formation of postoperative intestinal adhesion effectively.

Objective To evaluate the effect of diabetic peripheral neuropathy on peripheral neurotoxicity induced by local anesthetics in rats.Methods Sixty healthy adult male SPF Sprague-Dawley rats,aged 6 weeks,weighing 150-180 g,were divided into either control group (n =18) or diabetic peripheral neuropathy group (n=42) using a random number table.The rats were fed a high-fat and high-sucrose diet for 8 weeks,and streptozotocin (STZ) 30 mg/kg was injected intraperitoneally to induce diabetes mellitus which was confirmed by blood glucose level≥ 16.7 mmol/L.The mechanical paw withdrawal threshold to yon Frey filament stimulation and thermal paw withdrawal threshold were measured.The decrease in reaction thresholds to thermal and mechanical stimuli (changing from sensitivity to insensitivity) was observed after STZ injection.At 4 weeks after STZ injection,the rats showing a marked hyperalgesia served as early diabetic group.At 8 weeks after STZ injection,the rats showing a marked insensitivity to pain served as late diabetic group.Experiments were carried out in early or late diabetic rats,and ordinary Sprague-Dawley rats of the same age were used as control group.Left sciatic nerve block was performed with 2％ lidocaine 0.2 ml.Before the sciatic nerve block and at 1 week after the sciatic nerve block,the nerve conduction velocity of the left sciatic nerve and F-wave minimal latency were measured,and the sciatic nerve block time was recorded.Results Compared with the baseline before block,the nerve conduction velocity was significantly decreased,and the F-wave minimal latency was prolonged in late diabetic rats (P＜0.05).Compared with control group,the sciatic nerve block time was significantly prolonged in late diabetic group (P＜0.05).Conclusion Diabetic peripheral neuropathy aggravates peripheral neurotoxicity induced by local anesthetics in rats.

Objective:To evaluate the accuracy of two-dimensional ultrasound(2D-US), three-dimensional volume ultrasound (3D-US) and ultrasound strain elastography (USE) in the measurement of preoperative tumor size of breast cancer and its influencing factors.Methods:A total of 101 patients with breast cancer in Nanfang Hospital of Southern Medical University from April to November 2016 were recruited in this study. The maximum diameter of the lesion was examined by 2D-US 3D-US and USE before core needle biopsy or surgery biopsy. The Bland-Altman analysis and intraclass correlation coefficient (ICC) were used to analyze the consistency between the ultrasonic technique measurements and the pathological measurements of postoperative lesion. Chi-square test or Fisher exact test was used to analyze whether the accuracy of three imaging techniques was affected by different clinical pathologic factors and imaging characteristics.Results:3D-US showed better agreement with histology than 2D-US and USE, with a higher ICC (ICC 3D-US=0.90>ICC 2D-US=0.81>ICC SUE=0.78) and low variation. In 3D-US, the accuracy rate of the age >40 years old group was higher than ≤40 years old group. In 2D-US, the measurement accuracy of invasive ductal carcinoma (IDC) without intraductal carcinoma in situ (DCIS) group was higher than DCIS with DCIS group, non-microcalcifications group was more accurate than microcalcifications group. The long diameter of lesion ≤2 cm group was more accurate than >2 cm group, IDC group was more accurate than invasive lobular carcinoma(ILC) group. In USE, the measurement accuracy of IDC without DCIS group was higher than DCIS with DCIS group, non-microcalcifications group was more accurate than microcalcifications group. All the differences mentioned above were statistically significant(all P<0.05). Conclusions:For accurate measurement of the size of breast cancer lesions, 3D-US is the best, which is least affected by clinicopathological factors and imaging features, followed by 2D-US and USE. This has certain significance for clinically determining the extent of breast cancer lesions.

Objective To establish the methods of three-dimensional eonformal(3DCRT) and intensity-modulated radiotherapy(IMRT) for whole pelvic irradiation in post-hysterectomy cervical carcinoma, And to optimize the methods for clinical practice. Methods Between 2004 and 2005,10 patients with cervical carcinoma who underwent hysterectomy with high risk of recurrence were selected for this study. The following observations and measurements were used for the study: Set-up errors with supine or prone position were measured to determine appropriate immobilization position. Influence of full and empty bladder on irradiated normal tissue volume was measured. Treatment errors were detected and CTV/PTV were then delineated. 3DCRT and IMRT planning and comparison were applied. Results The set-up error was within 5 mm of three dimensions in prone position and more than 5 mm in supine position, the difference of which was statistically significant. The percentage of irradiated volume of the bladder and bowel was smaller when the bladder was full comparing with empty bladder. In prone position and with full bladder,portal films showed the movement of isocenter in three directions. The total uncertainty was [7.4±1.6]mm. For 95% confidence interval,the margin from CTV to PTV was 1 cm. CIPTV for 3,4,5,and 6 fields 3DCRT was 0.46,O. 67, O. 68, and O. 68, respectively. When beyond 4 fields, the advantage of adding fields was not significant.Four fields planning was feasible for clinical practice. CI for 5,7,9,11 ,and 13 fidds IMRT was 0.75,0.83, 0.84,0.85 ,and 0.85 ,respectively. When beyond 9 fields,the advantage of adding fields was not significant. Nine fields planning was feasible for clinical practice. Conclusions For whole pelvic radiotherapy for post-hysterectomy cervical carcinoma,prone position was better than supine position for immobilization due to smaller set-up errors. The full bladder is recommended during radiotherapy, planning,For clinical practice,4 fields planning is feasible in 3DCRT while 9 fields planning is feasible in IMRT.

Objective:To explore the effect of hydrogen sulfide (H 2S) on modulating the subunit Kir6.2 of adenosine triphosphate sensitive potassium channels via the cyclic guanosine monophosphate-dependent protein kinase (cGMP/PKG) signaling pathway in epileptic rat models. Methods:Sixty adult male SD rats were randomly divided into the following six groups (10 rats in each group) by random number table method: control, epileptic, H 2S donor, H 2S donor+epileptic, KT5823 (one inhibitor of the cyclic guanosine monophosphate-dependent protein kinase)+H 2S donor+epileptic, and glibenclamide (one inhibitor of the adenosine triphosphate sensitive potassium channels)+H 2S donor+epileptic groups. Except the control group, SD rats were intraperitoneally injected with plentylenetetrazole to make the kindling models and their behaviours were recorded including the latency period, the grade, and the duration of the first epileptic seizure according to the Racine′s standard. The waveforms of electroencephalogram (EEG) in hippocampus were also recorded during the seizure. The mRNA and protein levels of PKG and Kir6.2 in hippocampus were evaluated by Western blotting and quantitative real-time polymerase chain reaction, and the hippocampal concentrations of cGMP and phosphorylation of cyclic guanosine monophosphate-dependent protein kinase (p-PKG) were detected by enzyme linked immunosorbent assay. Results:Rats in the epileptic group showed Ⅳ-Ⅴ grade of epileptic seizure [4.500 (4.000, 4.875)], short latency period [(10.37±8.21) min] but long duration [(69.50±24.37) s] of seizure. Compared to the epileptic group, rats in the H 2S donor group showed Ⅱ-Ⅲ grade of epileptic seizure ( P=0.004), significantly longer latency period ( P<0.001), and shorter duration of seizure ( P<0.001). Compared to the H 2S donor+epileptic group, rats in the KT5823+H 2S donor+epileptic group showed Ⅲ-Ⅳ grade of epileptic seizures, significantly shorter latency period ( P<0.001), and longer duration of seizure ( P<0.001). The results of EEG showed that the wave patterns in the epileptic group were spike or sharp waves and the amplitudes were largest [(190.570±23.590) μV]. Compared with the epileptic group, amplitudes were reduced ( P<0.001) in the H 2S donor+epileptic group. PKG mRNA and PKG protein were expressed differently among all groups (PKG mRNA: n=5, H=26.714, P<0.001; PKG protein: n=5, F=30.597, P<0.001). Compared with the control group, the expression of both PKG mRNA and PKG protein was decreased (PKG mRNA: 1.000±0.001 vs 0.782±0.064, P=0.023; PKG protein: 0.550±0.037 vs 0.145±0.020, P=0.042) in the epileptic group. Besides, Kir6.2 mRNA and Kir6.2 protein were expressed differently among all groups (Kir6.2 mRNA: n=5, H=27.761, P<0.001; Kir6.2 protein: n=5, F=60.659, P<0.001). Compared with the control group, the expression of both Kir6.2 mRNA and Kir6.2 protein was decreased (Kir6.2 mRNA: 1.000±0.001 vs 0.897±0.033, P=0.004; Kir6.2 protein: 0.384±0.035 vs 0.215±0.016, P=0.024) in the epileptic group. And the concentrations of cGMP and p-PKG were decreased (cGMP: P<0.001; p-PKG: P<0.001) in the epileptic group. The results in the H 2S donor+epileptic group were up-regulated (PKG mRNA: P=0.047; PKG protein: P<0.001; Kir6.2 mRNA: P=0.011; Kir6.2 protein: P<0.001; cGMP: P<0.001; p-PKG: P<0.001) compared with the epileptic group. However, the results in the KT5823+H 2S donor+epileptic group were down-regulated (PKG mRNA: P=0.015; PKG protein: P=0.027; Kir6.2 mRNA: P=0.013; Kir6.2 protein: P=0.017; cGMP: P=0.005; p-PKG: P<0.001) compared with the H 2S donor+epileptic group. Conclusion:A possible mechanism is that H 2S prevents the epileptic seizure from modulating the subunit Kir6.2 of ATP sensitive potassium channels via the cGMP/PKG signaling pathway.

Objective To evaluate the role of NOX2 in bupivacaine-induced production of reactive oxygen species (ROS) in nerve cells.Methods SH-SY5Y cells were seeded in culture plates and divided into 4 groups (n =11 each) using a random number table:small interfering RNA (siRNA) negative control group (group NC),siRNA negative control plus bupivacaine group (group NC +B),NOX2 siRNA group and NOX2 siRNA plus bupivacaine group (group NOX2 siRNA + B).In NC and NOX2 siRNA groups,the cells were transfected with negative siRNA and NOX2 siRNA,respectively,and then incubated in the culture medium for 24 h.In NC+B and NOX2 siRNA+B groups,cells were transfected with negative siRNA and NOX2 siRNA,respectively,new plates were used,the cells were incubated for 3 h with bupivacaine at the final concentration of 1.5 mmol/L,the culture medium was then replaced,and the cells were incubated until 24 h.The level of intracellular ROS was measured using the fluorogenic probe dihydroethidium,the cell apoptosis was determined by TUNEL,and the expression of activated caspase-3 and caspase-9 was detected using Western blot.Apoptosis rate was calculated.Results Compared with group NC,the level of ROS and apoptosis rate were significantly increased,and the expression of activated caspase-3 and caspase-9 was up-regulated in group NC+B (P＜ 0.05),the level of ROS was significantly increased,and the expression of activated caspase-3 and caspase-9 was up-regulated (P＜0.05),and no significant change was found in apoptosis rate in group NOX2 siRNA+B (P＞0.05),and no significant change was found in the level of ROS or apoptosis rate (P＞0.05),and the expression of activated caspase-3 and caspase-9 was significantly up-regulated in group NOX2 siRNA (P＜ 0.05).Compared with group NC+B,the level of ROS and apoptosis rate were significantly decreased,and the expression of activated caspase-3 and caspase-9 was down-regulated in group NOX2 siRNA+B (P＜0.05).Conclusion NOX2 is involved in the pathophysiological mechanism of bupivacaine-induced burst production of ROS in nerve cells.