1.Mechanisms of tumor necrosis factor-related apoptosis inducing ligandcombined with Triptolide in inducing the apoptosis of pancreatic cancer cells
Yan JIANG ; Wei LIU ; Zhiyu CHEN
Chinese Journal of Digestive Surgery 2015;14(4):321-328
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.
2.Diagnosis and treatment of the sphenoid sinus malignant tumor
Xing YE ; Zhiyu YAN ; Guangliang GUO ;
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(09):-
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.
3.Doppler ultrasound screening for deep vein thrombosis in patients with lower limb fracture
Youqing YAN ; Hua REN ; Kun FENG ; Chaohui DU ; Zhiyu WEI
Chinese Journal of General Practitioners 2011;10(9):673-674
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.
4.Three-dimensional printing technology-aided total knee arthroplasty for osteoarthritis with genu varum deformity
Zhenguo SUN ; Jiajun ZHU ; Yan CUI ; Shenghui NI ; Zhiyu ZHANG
Chinese Journal of Tissue Engineering Research 2017;21(15):2395-2399
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.
5.Insulin-like growth factor 1 promotes in vitro proliferation of chondrocytes in traumatic arthritis
Guang ZHAO ; Jifeng JING ; Zhiyu ZHANG ; Jiajun ZHU ; Yan CUI
Chinese Journal of Tissue Engineering Research 2014;(2):183-186
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.
7.Research on Digitalization Path and Method of Hospital Medical Records Information
Xiangpei WANG ; Yipeng SONG ; Liyun HE ; Zhiyu WANG ; Dongning YAN ; Yang YANG ; Peng LI
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(2):389-393
Based on a retrospective study design, this article was aimed to discuss the transformation path and method of hospital medical records information into analyzable data, in order to solve the problem of text-based information digitalization, and improve the credibility of statistical results. The hospital medical records information of traditional Chinese medicine (TCM) treatment for infantile cerebral palsy was taken as example. After the identification of research purpose, the study contained 8 steps, which were index identification, index definition, coding and assignment, database design, data entry, quality control, and database locking. The database contained research-related indicators was received. The results showed that a set of path and method based on indicator extraction and conversion of hospital medical records information into data. It was concluded that in the retrospective study design, the conversion of text information into analyzable data was the key step. Extraction and digitalization of indicators should be based on the research plan. Key elements such as the crowd, treatment plan, evaluation indicator, should be paid attention to for the systematic analysis, in order to ensure the system integrity of research indicators.
8.Ischemic postconditioning attenuates myocardial cell injury by ischemia-reperfusion injury in rabbits
Yan HE ; Zhiyu ZENG ; Jinyi LI ; Guoqiang ZHONG ; Wei LI ; Weike LI ; Honghong KE
Basic & Clinical Medicine 2010;30(2):133-138
Objective To investigate the effects of ischemic postconditioning on apoptosis, structural and functional changes of mitochondria induced by myocardial isehemia/reperfusion (I/R) injury of rabbits and potential mechanism. Methods Eighty healthy rabbits were divided randomly into five groups: sham operation group ( Group Sham) , ischemic reperfusion group (Group IR) , ischemic preconditioning group (Group IP) , ischemic postconditioning group (Group PC) and 5-HD plus ischemic postconditioning group (Group PC +5-HD). All rabbits in the five groups were killed 4 h after reperfusion. The hearts were quickly collected for microscopy by TUNEL. We observed ultrastructural changes of myocardium under electron microscope and examined mitochondrial membrane potential and Ca~(2+) concentration, MDA content and SOD activity of myocardial mitochondria. Results Compared with group IR, the damage of mitoehondrial ultrastrueture was milder, the apoptosis rate decreased and Ca concentration and MDA content were much lower in group IP and group PC ( P < 0. 05 ). Mitochondrial membrane potential and SOD activity of myocardial mitochondria in group IP and group PC was significantly higher than that in group IR(P<0.05). The protective effect of PC against I/R injury was partially counteracted by 5-HD .Conclusion Ischemic posteonditioning can protect the heart from I/R injury, this is supported by improvement mitochondrial ultrastructure and by decreasing apoptosis, increasing mitochondrial membrane potential and SOD activity, alleviating Ca~(2+) overload and decreasing MDA content in myocardial mitochondria. The cardio protective effects may be explained by mitochondrial ATP sensitive potassium channel.
9.Three-dimensional total endoscopic thyroidectomy for thyroid carcinoma: a series of 32 cases.
Yong WANG ; Qunzi ZHAO ; Haichao YAN ; Qiuping XIE ; Zhiyu LI ; Ping WANG ; Shaoming XU
Chinese Journal of Surgery 2015;53(3):176-178
OBJECTIVETo summarize the experiences of applying three-dimensional (3D) technique in scarless endoscopic thyroidectomy (SET) via anterior chest approach.
METHODSMedical records of patients who undenvent SET using 3D technique from December 2013 to May 2014 were retrospectively reviewed. A total of 32 patients who had a preoperative ultrasound-guided fine-needle aspiration cytology diagnosis of unilateral papillary thyroid carcinoma and underwent lobectomy associated with central lymph node dissection were included in this study. All patients were female with a mean age of (37 ± 10) years at diagnosis. In addition, 45 female patients who underwent traditional endoscopic thyroidectomy at the same period were randomly selected as control.
RESULTSAll surgical procedures were successfully finished. The mean surgical time in 3D group was (91. 7 ± 11. 4) minutes, and mean hospitalization time was (3. 2 ± 0. 5) days. Contemporary hoarseness was observed in one patient, and no bleeding, infection, hypocalcemia and other postsurgical complications were observed. Compared to the traditional endoscopic surgery group, 3 D group had significantly shorter surgical time of lobectomy ((23. 2 ± 5. 1) minutes vs. (28. 0 ± 5. 0) minutes, t = 4. 156, P = 0. 000). Negative results were seen in the time taking of creating surgical space ((14. 6 ± 3. 3) minutes vs. (15. 6 ± 2. 5) minutes), central lymph node dissection ((25. 1 ± 5. 4) minutes vs. (24. 4 ± 6. 3) minutes) and total surgical time ((91.7 ± 11. 4) minutes vs. (96. 1 ± 13. 0) minutes).
CONCLUSIONApplication of 3D technique in endoscopic surgery can offer 3D vision of the surgical field, thus significantly shorten lobectomy time and more easily to achieve fine dissection and functional protection of recurrent laryngeal nerves, parathyroids and other vital anatomic structures.
Adult ; Biopsy, Fine-Needle ; Carcinoma ; diagnosis ; surgery ; Carcinoma, Papillary ; Endoscopy ; methods ; Female ; Humans ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Operative Time ; Retrospective Studies ; Thyroid Neoplasms ; diagnosis ; surgery ; Thyroidectomy ; methods
10.Evaluation of left ventricular systolic transmural myocardial dysfunction of maintenance hemodialysis patients using ultrasonic layer‐specific strain technology
Shuangshuang YAN ; Lixue YIN ; Hongmei ZHANG ; Yanping CHEN ; Zhiyu GUO
Chinese Journal of Ultrasonography 2019;28(3):205-210
Objective To assess the clinical ultrasound value of layer‐specific strain in evaluation of left ventricular systolic myocardial dysfunction of uremia patients after long‐time dialysis at different time . Methods A total of 68 uremia patients accepted maintenance hemodialysis ( M HD ) were enrolled . T he patients were divided into two groups according to the dialysis duration :dialysis time <3 years ( group B , n=31) and dialysis time ≥3 years ( group C , n =37) . T he age and sex mached healthy cases were selected as control group ( group A , n = 30 ) . T he standard dynamic two‐dimensional echocardiographic viewes of apical four‐chamber ,three‐chamber ,two‐chamber and the short‐axis view at three levels of mitral valve , papillary muscle and apex were acquired for three cardiac cycles . T he highest value of peak systolic longitudinal strain ( LS ) ,circumferential strain ( CS ) at different levels ,left venrticular global longitudinal strain ( GLS ) and global circumferential strain ( GCS ) were respectively assessed from endocardium ,mid‐myocardium and epicardium using GE EchoPAC workstation . T he comparisons of those parameters were performed among the 3 groups for differences . T he efficacies of GLS and GCS at different myocardial layers in diagdosing the left ventricular systolic function of M HD patients were analyzed by the ROC curve . Results ① Global transmural parameters :compared with those in group A ,the values of GLS at three myocardial layers in both M HD groups were significantly decreased ( all P < 0 .01 ) ,the value of GLS at three myocardial layers in group C was also decreased ,and was statistically different from that in group B ( P<0 .01) . Compared with those in group A ,the values of GCS at mid‐myocardium in group B and three myocardial layers in group C were also decreased ( all P <0 .01) . T here was no significant difference of GCS between group B and C ( P >0 .05) . ②Longitudinal transmural parameters at different levels :the values of LS at three myocardial layers of mitral valve ,papillary muscle and apex were decreased in group B and C compared with those in group A ( P <0 .05 or P <0 .01) ; T he values of LS at three myocardial layers of mitral valve ,papillary muscle and apical levels were also decreased in group C compared with those in group B ( P <0 .05 or P <0 .001) . ③Short‐axis transmural parameters at different levels :compared with those in group A ,the value of CS at mid‐myocardium of mitral valve level was decreased in group B ( P <0 .05) ,the values of CS at three myocardial layers of the mitral valve level and mid‐myocardium of papillary muscle level and apical level were decreased in group C ( P <0 .05 or P <0 .01) . Besides ,compared with those in group B ,the values of CS at mid‐myocardium and epicardium of mitral valve level were also decreased in group C ( P <0 .05) . ④ROC curve showed that determining left ventricular systolic dysfunction in M HD patients using GLS ,GCS at different myocardial layers ,when the area under the curve ( AUC ) of GLS of intima was 0 .851 ,the cut‐off value was -21 .45% ,the sensitivity was 72 .7% ,and the specificity was 93 .3% ; when the AUC of GCS of mid‐myocardium was 0 .683 ,the cut‐off value was -17 .08% , the specificity was 58 .5% , and the specificity was 83 .3% . Conclusions T he left ventricular systolic myocardial function is progressively damaged with the extended dialysis duration time . Ultrasonic layer‐specific strain technology could be used to quantitatively evaluate left ventricular systolic transmural myocardial dysfunction and might contribute to the evaluation of the severity of left ventricular myocardial dysfunction clinically for a more accurate intervention .