Objective To investigate the mechanisms of tumor necrosis factor-related apoptosis inducing ligand (TRAIL) combined with Triptolide in inducing the apoptosis of pancreatic cancer cells.Methods (1) The pancreatic cancer cells (MiaPaca-2 cells) were divided into 4 groups:blank control group (no drugs were added),TRAIL + Triptolide-group (only TRAIL was added),TRAIL-Triptolide + group (only Triptolide was added) and TRAIL+ Triptolide+ group (TRAIL and Triptolide were added).The vitality of cells in all the 4 groups was assessed by CCK-8.The expressions of poly ADP-ribose polymerase (PARP),cysteinyl aspartate specific proteinase-3 (Caspase-3) and Caspase-8 were detected by Western blot.The vitality of cells was detected by CCK-8 and the vitality of Caspase-8 was detected by Caspase-Glo assays after adding Z-IETD-FMK,a specific inhibitor of Caspase-8.The expressions of myeloid cell leukemia-1 (Mcl-1),Bcl-xL and Bcl-2 were detected by Western blot.(2) The MiaPaca-2 cells were divided into 8 groups:①TRAIL-Mcl-1 siRNA-group (no TRAIL was added and Mcl-1 siRNA cells were not transfected),TRAIL+ Mcl-1 siRNA-group (TRAIL was added and Mcl-1 siRNA cells were not transfected),TRAIL-Mcl-1 siRNA + group (TRAIL was not added and Mcl-1 siRNA cells were transfected)and TRAIL+ Mcl-1 siRNA+ group (TRAIL was added and Mcl-1 siRNA cells were transfected).②TRAIL-Bcl-xL siRNA-group (TRAIL was not added and Bcl-xL siRNA was not transfected),TRAIL+ Bcl-xL siRNA-group (TRAIL was added and Bcl-xL siRNA was not transfected),TRAIL-Bcl-xL siRNA + group (TRAIL was not added and Bcl-xL siRNA was transfected) and TRAIL+ Bcl-xL siRNA+ group (TRAIL was added and Bcl-xL siRNA was transfected).The vitality of the cells in all the groups was detected by CCK-8.The expressions of Caspase-3 and Caspase-8 protein were detected by Western blot.The measurement data with normal distribution were presented as (x) ± s.The comparison among groups was done by ANOVA,and the pairwise comparison was done by LSD-t test.Results (1) The vitalities of MiaPaca-2 cells in the blank control group,TRAIL + Triptolide-group,TRAIL-Triptolide + group and TRAIL + Triptolide + group were 100.0％ ± 1.1％,81.2％ ± 2.3％,78.6％ ± 3.6％and 40.1％ ± 2.5 ％,and the relative expressions of PARP protein were 0.510 ± 0.028,0.720 ±0.072,1.250 ±0.023 and 2.560 ± 0.220,the relative expressions of Caspase-3 were 0.080 ± 0.004,0.080 ± 0.003,0.110 ±0.005 and 2.720 ± 0.003,and the relative expressions of Caspase-8 were 0.070 ± 0.003,0.080 ± 0.005,0.120 ±0.003 and 0.990 ± 0.006,with significant differences among the 4 groups (F =203.607,1 457.785,332 421.900,35 437.218,P ＜ 0.05).The vitality of M iaPaca-2 cells in the TRAIL + Triptolide + group was significantly different from those in the blank control group,the TRAIL + Triptolide-group and the TRAIL-Triptolide + group (t =34.583,355.936,36.271,P ＜ 0.05).The relative expression of PARP protein of MiaPaca-2 cells in the TRAIL+ Triptolide + group was significantly different from those in the blank control group,TRAIL+ Triptolidegroup and TRAIL-Triptolide + group (t =591.784,63.739,2 268.987,P ＜ 0.05).The relative expression of Caspase-3 protein of the MiaPaca-2 cells in the TRAIL + Triptolide + group was significantly different from those in the blank control group,the TRAIL + Triptolide-group and theTRAIL-Triptolide + group (t =3 266.153,9 145.228,1 738.713,P ＜0.05).The relative expression of Caspase-8 protein of the MiaPaca-2 cells in the TRAIL+ Triptolide +group was significantly different from those in the blank control group,the TRAIL+ Triptolide-group and the TRAIL-Triptolide + group (t =663.953,l 432.878,327.584,P ＜ 0.05).The vitality of caspase-8 in the TRAIL+ Triptolide+ group was 711.0％ ± 5.1％ before adding Z-IETD-FMK,and then the vitality of MiaPaca-2 cells and caspase-8 changed to 70.0％ ± 4.8％ and 73.0％ ± 2.4％,with significant differences (t =17.956,55.027,P ＜ 0.05).The relative expressions of Mcl-1 protein in the blank control group,the TRAIL + Triptolidegroup,the TRAIL Triptolide + group and the TRAIL + Triptolide + group were 1.68 ± 0.22,2.08 ± 0.11,0.73 ±0.15 and 0.58 ± 0.18,the relative expressions of Bcl-xL protein were 0.65 ± 0.03,0.47 ± 0.03,0.32 ± 0.03and 0.26 ±0.05,the relative expressions of Bcl-2 protein were 0.65 ± 0.03,0.67 ± 0.03,0.62 ± 0.05 and 0.67 ± 0.03,with significant difference among the 4 groups (F =55.178,88.683,3.411,P ＜ 0.05).The relative expressions of Mcl-1 protein of the MiaPaca-2 cells in the TRAIL-Triptolide + group and the TRAIL+ Triptolide +group were significantly different from those of the blank control group (t =23.506,47.631,P ＜ 0.05) and the TRAIL + Triptolide-group (t =58.457,37.115,P ＜ 0.05).The relative expressions of Bcl-xL protein of the MiaPaca-2 cells in the TRAIL-Triptolide + group and the TRAIL + Triptolide + group were significantly different from those of the blank control group (t =38.105,42.219,P ＜ 0.05) and the TRAIL + Triptolide-group (t =32.476,15.814,P ＜ 0.05).The relative expressions of Bcl-2 protein in the TRAIL-Triptolide + group and the TRAIL+ Triptolide + group were not significantly different from those of the blank control group (t =4.724,1.732,P > 0.05) and the TRAIL + Triptolide-group (t =3.464,0.000,P ＞ 0.05).(2) The vitalities of MiaPaca-2 cells of the TRAIL-Mcl-1 siRNA-group,TRAIL + Mcl-1 siRNA-group,the TRAIL-Mcl-1 siRNA + group and the TRAIL + Mcl-1 siRNA + group were 100.0％ ± 2.2％,79.3％ ± 1.8％,71.2％ ± 3.2％ and 37.3％ ± 5.4％,the relative expressions of Caspase-8 protein were 0.100 ± 0.003,0.100 ± 0.005,0.100 ± 0.003 and 0.350 ±0.005,and the relative expressions of Caspase-3 protein were 0.020 ± 0.003,0.060 ± 0.003,0.020 ± 0.003 and 0.590 ±0.004,with significant differences among the 4 groups (F =136.681,2 717.391,44 471.429,P ＜0.05).The vitality of MiaPaca-2 cells of the TRAIL + Mcl-1 siRNA + group was significantly different from those in the TRAIL-Mcl-1 siRNA-group,the TRAIL + Mcl-1 siRNA-group and the TRAIL-Mcl-1 siRNA + group (t =33.937,20.207,26.689,P ＜ 0.05).The relative expression of Caspase-8 protein of the TRAIL + Mcl-1 siRNA +group was significantly different from those in the TRAIL-Mcl-1 siRNA-group,the TRAIL + Mcl-1 siRNA-group and the TRAIL-Mcl-1 siRNA + group (t =216.506,433.013,144.338,P ＜ 0.05).The relative expression of Caspase-3 protein of the TRAIL + Mcl-1 siRNA + group was significantly different from those in the TRAIL-Mcl-1 siRNA-group,the TRAIL + Mcl-1 siRNA-group and the TRAIL-Mcl-1 siRNA + group (t =329.09,458.993,987.269,P ＜0.05).The vitalities of MiaPaca-2 cells of the TRAIL-Bcl-xL siRNA-group,the TRAIL+ Bcl-xL siRNA-group,the TRAIL-Bcl-xL siRNA + group and the TRAIL+ Bcl-xL siRNA + group were 100.0％ ± 2.3％,87.2％ ± 4.1％,74.1 ± 3.7％ and 56.3％ ± 5.4％,and the relative expressions of Caspase-3 protein were 0.060 ±0.004,0.070 ± 0.003,0.060 ± 0.004 and 0.390 ± 0.003,with significant differences among the 4 groups (F =70.074,4 643.478,P ＜ 0.05).The vitality of MiaPaca-2 cells of the TRAIL + Bcl-xL siRNA + group was significantly different from those in the TRAIL-Bcl-xL siRNA-group,the TRAIL+ Bcl-xL siRNA-group and the TRAIL-Bcl-xL siRNA + group (t =24.416,41.170,18.136,P ＜ 0.05).The relative expression of Caspase-3 protein of the TRAIL + Bcl-xL siRNA + group was significantly different from those in the TRAIL-Mcl-1 siRNA-group,the TRAIL + Mcl-1 siRNA-group and the TRAIL-Mcl-1 siRNA + group (t =285.788,554.256,190.526,P ＜ 0.05).Conclusion Triptolide could induce the apoptosis of MiaPaca-2 cells by inhibiting the expressions of Mcl-1 and Bcl-xL,sensitizing TRAIL and activating Caspase-8 and Caspase-3.

OBJECTIVE To investigate the clinical characteristics,nasal endoscope and imaging findings, misdiagnosis and treatment results of sphenoid sinus malignant tumor.METHODS The clinical data of 9 cases with sphenoid sinus malignant tumor were summarized and analyzed.Headache was found in 8 patients, ophthalmic symptoms in 3 patients,nasal bleeding and obstruction in 3 patients,and cranial nerve palsy in 2 patients.They were often misdiagnosed as sphenoid sinusitis and nasopharyngeal carcinoma.RESULTS Only 1 patient with papillary carcinoma was cured for 6 years.One patient with neuroendocrine carcinoma was alive for 3 years after treatment.Four patients died at 2 to 3 years.Two patients were alive at 1 year after operation. One patient was lost to follow up.CONCLUSION Sphenoid sinus tumor had no characteristic symptoms in early stage.It is easy to misdiagnose and delay diagnosis. Patients with headache,visual symptoms and nasal bleeding should take nasal endoscopy,CT scan and MRI examination at early stage.

BACKGROUND: The complicated localization of intramedullary nails and osteotomy more dependent on surgeons' experience limit the application of conventional total knee arthroplasty (TKA). The occurrence of three-dimensional (3D) printing technology can achieve precise localization and osteotomy in TKA.OBJECTIVE: To explore the effectiveness of 3D printing technology-aided TKA versus conventional TKA for genu varum.METHODS: Thirty-four patients with genu varum undergoing primary unilateral TKA were recruited and were then divided into two groups (n=17 per group) in accordance with the random number table. One group was treated with TKA with 3D printing guild plate (3D printing group), while the other group received the conventional TKA (conventional group).The intraoperative and postoperative blood loss, operation time, as well as the Hospital for Special Surgery score, range of motion, and lower limb mechanical alignment at 2 weeks postoperatively were compared between two groups.RESULTS AND CONCLUSION: (1) The range of motion of knee in the 3D printing group was larger than that in the conventional group, but had no significant difference at 2 weeks postoperatively (P=0.744). (2) There was no significant difference in the Hospital for Special Surgery scores between two groups at 2 weeks postoperatively (P= 0.532). (3) The postoperative lower limb mechanical alignment showed no significant difference between two groups (t=0.218, P=0.632).(4) The operation time in the 3D printing group was significantly shorter than that in the conventional group (P=0.000). (5) The blood loss in the 3D printing group was significantly less than that in the conventional group (P=0.000). (6) Our findings indicate that 3D printing technology-aided TKA exhibits similar results to the conventional TKA in the Hospital for Special Surgery scores, range of motion, and lower limb mechanical alignment, but it shortens the operation time,reduces the blood loss, and achieves precise osteotomy, which is available for the elderly with poor basic condition, and weak tolerance of surgery.

One hundred and sixteen patients with lower limb fracture were screened by Doppler ultrasound for deep vein thrombosis(DVT) of bilateral lower extremities within the first 72 h, d7 and d21 after fracture. Results showed that DVT was detected in 31 (26. 7% ) out of 116 cases within 72 h; at d7 and d21 DVT was detected in 3 and 1 patient respectively with a cumulated DVT rate of 30. 2% in 3 weeks.Serial Doppler ultrasonography is of value in screening for DVT of the lower extremities in patients with lower limb fracture at early stage.

BACKGROUND:The early damaged chondrocytes are susceptible to de-differentiate and exert unstable phenotype during the in vitro culture, thus needing some growth factors.
OBJECTIVE:To observe the promotion effect of insulin-like growth factor 1 on the in vitro proliferation of chondrocytes in adult rabbits with traumatic arthritis.
METHODS:Traumatic arthritis models of adult rabbits were established by using the modified Hulth method. After the models were successful y established, the distal femur and proximal tibia were harvested under sterile conditions, the chondrocytes were cultured. The cultured cells were divided into two groups:control group was cultured with Dulbecco’s modified Eagle’s medium containing 10%fetal bovine serum, while experimental group was cultured with Dulbecco's modified Eagle’s medium containing 100μg/L insulin-like growth factor 1. The effect of insulin-like growth factor 1 on the proliferation of chondrocytes in adult rabbits with traumatic arthritis was determined through the cytomorphology, cellcounting, and cellactivity.
RESULTS AND CONCLUSION:The chondrocytes in adult rabbits with traumatic arthritis were successful y cultured, the majority of cells were mini-cells, presenting smal fusiform, round or polygonal shape. Hematoxylin-eosin staining showed that the number of cells in experimental group was higher than that in control group. MTT assay found that the absorbance of cells in experimental group was greater than that in control group (P<0.01). Our findings indicate that, insulin-like growth factor 1 can promote the in vitro proliferation of chondrocytes in adult rabbits with traumatic arthritis.

Objective To assess the left ventricular systolic function after renal sympathetic denervation(RSD) in beagles with heart failure by layer-specific two-dimensional speckle tracking imaging.Methods Twenty-three beagles were randomly divided into control group (8 dogs),URSD group (8 dogs)and RSD group (7 dogs).Control group was implanted with pacemakers without pacing,URSD group was implanted with pacemakers and underwent 3 weeks of rapid right ventricular pacing,and RSD group underwent catheter-based RSD bilaterally and was simultaneously implanted with pacemakers.All standard measurements were obtained from the left ventricular short-axis views at the levels of mitral annulus,papillary muscle and apex,and apical four-chamber,two-chamber,and long-axis views.The peak systolic longitudinal strain and circumferential strain of each layer of the left ventricle were measured by layerspecific two-dimensional speckle tracking imaging.Results After 3 weeks,all of the dogs in both the URSD and RSD groups showed lower left ventricular diastolic and systolic function compared with those in the control group,however,the dogs in the RSD group had a higher LV ejection fraction (LVEF) than the dogs in the URSD group(P ＜ 0.05).Compared with the control group,the global longitudinal strain,circumferential strain and radial strain all decreased significantly in URSD and RSD groups.The longitudinal strain of endo-and mid-cardium and the circumferential strain in endo-cardium were significantly higher in RSD group than those in URSD group(P ＜0.05).Conclusions Myocardial systolicfunction is impaired in dogs with heart failure,layer-specific two-dimensional speckle tracking imaging can provide effective tools to assess the left ventricular systolic function.Renal sympathetic denervation can improve the myocardial systolic function in dogs with heart failure.

Objective To compare differences and similarities of the content of trigonelline in Himalaica mirabilis of either wild or cultivated materials from different places. Methods An HPLC method was established to determine the content of trigonelline in Himalaica mirabilis. ZORBAX XDB-CN column (4.6 mm×250 mm, 5 μm) was used, with mobile phase of acetonitrile-0.03%acetic acid (85∶15), flow rate of 0.8 mL/min, detection wavelength of 265 nm, determination wavelength of 360 nm, and column temperature of 30 ℃. Results Regression calculation was made on peak area with the reference solution concentration, and then got the regression equation A=23.409C-26.398, r=0.999 8. Trigonelline showed good linear relation with peak area among the range of 2.004-200.400 μg/mL. The average recovery of trigonelline was 99.57%, RSD=1.11%. Conclusion There was no significant difference in the content of trigonelline of either wild or cultivated materials from different places. This study laid the foundation of application of the cultivated Himalaica mirabilis.

Objective To study on the association of apolipoprotein E(apoE)genotype with coronary heart disease(CHD)and type 2 diabetes mellitus(T2DM).Methods PCR-RFL,multiplex amplification refractory mutation system(muli-ARMS)and PCR-SSCP methods were used to detect the genotype of apoE,and DNA sequencing technique were used for further confrm the genotype and gene variations in 2 446 Chinese individuals,including 238 cases of CHD,316 cases of T2DM and 1 892 healthy controls.Fasting blood glucose(FBG)and plasma lipids levels[TC,TG,HDL-C,LDL-C,apoA I,apoB and Lo(a)]were measured by usual methods.Results Compared with the controls,plasma HDL-C(t=2.66)and apoA I(t=2.30)levels in the CHD group were significantly lower(P<0.05),but not in T2DM group;plasma TC level(t=5.22)in the T2DM group were significantly higher(P<0.05),but not in CHD group;systolic pressure(t=8.48,5.74)diastolic pressure(t=5.66,3.35),plasma TG(t=3.38, 4.56),LDL-C(t=2.48,7.00),apoB(t=1.67,2.24),Lp(a)(t=4.16,4.15)and FBG(t=7.04, 16.93)levels were significantly higher in both CHD group and T2DM group(P<0.05).The distributions of apoE ε2/2,ε2/3,ε3/3,ε2/4,ε3/4 and ε4/4 respectively were 0.4%,13.4%,58.0%,1.3%, 26.5%,0.4% in the CHD group;0.6%,5.7%,72.8%,1.9%,14.9%,4.1% in the T2DM group; 0.5%,10.5%,69.6%,1.6%,16.8%,1.1% in the control group.Significant differences were found between the CHD group(χ2=14.90,P=0.00),T2DM group(χ2=7.08,P=0.03)and the control group for the frequencies of apoE genotype.The distribution of ε3/4 was higher(26.5% vs 16.8%)and ε3/3 Was lower(58.0% vs 69.6%)in the CHD group.In the T2DM group.the distribution of εε4/4 Was higher (4.1% vs 1.1%),and 2 cases of ε3/3 with Arg 150 His mutation in exon 4 of apoE gene were firstly reposed in China,which is none in the CHD and control groups.Conclusions The results suggested that apoE ε3/4 and ε4 genotypes might be associated with the susceptibility of CHD and T2DM.respectively. To some extent,apoE ε3/3 may not be a good genotype for T2DM because of the Arg 150 His mutation. Blood pressure and plasma lipids could be used for diagnosis of the two diseases.

Objective To study the variation and characteristics of HIV-1 tat exon 1 gene from a patient with AIDS dementia complex( ADC), so as to research the pathogenesis of ADC. Methods The tat gene was amplified with nested PCR from genomic DNA which was extracted from lymph node, spleen and different brain tissues( meninges, grey matter from frontal cortex, white matter from frontal cortex, temporal cortex and basal ganglia) of a patient who died of ADC. PCR products were cloned into the pGEM-T vector,after transformation and selection by ampicillin and blue/white spotting. Five of positive clones were sequenced. HIV-1 tat sequences were processed with BioEdit and MEGA4. With the softwares, Neighbor-Joining tree, p-Distances, values of ds/dn, and analysis of amino acid motifs were all done. Results The samples were all identified as HIV-1 B and genetic variation exists in HIV-1 tat isolated from different tissue;Compared with HXB2, sixteen sites of the amino acid seque nce coded by the HIV-1 tat gene which was isolated from the patient changed. In addition, part of the changes were different between periphery and brain,especially, the five Q54R changes from basal ganglia and one Q54R change from temporal cortex are deserve to follow with interest. Conclusion Variations exist in the HIV-1 tat genes extracted from the ADC patient and the variations from peripheral and central nerve tissues were different, whether the variations concerned with the pathogenesis of ADC need more research.

Aim To investigate the roles of Cx43 in mitochondria and mitochondrial ATP sensitive potassium channe1(mitoK_(ATP)~+)participating in the protection for heptanol preconditioning on myocardial ischemia/reperfusion injury of rabbits.Methods In anesthetized open-chest rabbits,the left anterior descending artery(LAD)was occluded for 30 min and reperfused for 4 hrs.All rabbits were randomly divided into five groups(n=16 in each group):sham operation group(Group Sham),ischemic reperfusion group(Group IR),ischemic preconditioning group(Group IP),heptanol preconditioning group(Group HT)and 5-HD plus heptanol preconditioning group(Group HT+5-HD),All rabbits in the five groups were killed 4h after reperfusion.Myocardial infarct size was determined at the end of the experiment.The heart rate and the mean arterial pressure(MAP)were recorded and plasma CK-MB and cTnI activity were measured at baseline,the end of ischemia,and after 2 hrs and 4 hrs of reperfusion respectively.Mitochondria was isolated with different centrifugations.Ultrastructural changes of mitochondria were observed under electron microscope.The content of the mitochondria Cx43 was detected with Western blot.Results The plasma CK-MB,cTnI activity and myocardial infarct size were significantly reduced in IP(18.97±2.8)% and HT(19.97±3.8)% groups as compared to IR groups (35.67±5.8)%,(P<0.01).Compared to group IR,the damage of mitochondria in group IP and group HT were milder(P<0.01).No significant difference was found between Group IP and Group HT.Compared to sham group, the mitochondria Cx43 expression was distinctly decreased in group IR and group HT+5-HD(P<0.01)and no significant difference was found between Group IP and Group HT.Conclusions Heptanol preconditioning can protect the heart from I/R injury by improving mitochondrial ultrastructure and by attenuating the decrease of mitochondria Cx43 expression induced by I/R.The mitochondrial Cx43 expression may be concerned with depending on mito K_(ATP)~+.