1.THE CONSTRUCTION OF TRANSIENT GENE EXPRESSION TRANSFECTION SYSTEM IN MCF-7 CELLS
Haijiang ZHANG ; Shiwei HAN ; Jianli SANG ; Dacheng HE
Acta Anatomica Sinica 1954;0(02):-
Objective To establish an efficient, simple, low cytotoxicity and cheap transfaction system. Methods We have used the cationic polymer polyethylenimine(PEI) to study transient transfection in MCF-7 cells by testing different conditions, including cell concentrations, DNA concentrations, the ratio of PEI nitrogen to DNA phosphate(PEI-N∶DNA-P) and the time of cells grown in serum-free culture together with PEI-DNA complex. Results The optimized cell concentrations were 2?10 5 cells seeded per well in 24-well dishes 18-24?h before transfection. The DNA concentrations and ratio of PEI-N∶DNA-P are very important for optimal transfection and they affect each other. For 1??g DNA per well, the optimal PEI-N∶DNA-P is about 33∶1, however, as for 4??g DNA, it is 9∶1. The best time of cells grown in serum-free culture together with PEI-DNA complex is about 5-7?h.Conclusion With optimized conditions, we can establish an efficient, simple, low cytotoxicity and cheap transfection system by using PEI.
2.Clinical outcomes of operations in humeral intercondylar fractures
Yanhuang WANG ; Dacheng HAN ; Ming YANG ; Yongqing YAN ; Peixun ZHANG ; Zhongguo FU ; Hongbo ZHANG ; Dianying ZHANG
Clinical Medicine of China 2011;27(1):1-4
Objective To assess the efficienty of operation in humeral intercondylar fractures by section, reposition, internal fixation by composition nail and replacement of elbow joint. Methods From January, 1999 to May, 2009, Forty-six cases of the humeral intercondylar fracture were treated with operation. Thirty-eight cases were followed for 17 months. Their affected elbow joint function were evaluated by Cassebaum rating system, patients treated with total elbow replacement evaluated by Mayo elbow score, DASH score additionally. Results According to Cassebaum rating system,there were 22 cases rated as excellent,eight cases rated as good,five cases rated as poor,three case rated as poor. The fineness rate is 78. 9% (30/38). Mayo score in patients with joint replacement ranged from 75.0 - 90. 0, averaged 84. 4 ± 1.7. DASH score ranged from 25.0 to 75.0,averaged 41. 1 ±0. 8. There was 1 case of superficial soft tissue nonhealing and 2 cases of ulna nerve symptoms. Myodynamia in elbow joint bend and stretch was Ⅳ in 1 cases. Conclusion It is a good method to treat the humeral intercondylar fracture with rational use of open reduction or elbow replacement according to fracture type and patient condition.
3.Gene expression profile in osteoblastic differentiation of bone marrow stromal cells stimulated by simvastatin Gene chip analysis
Yaqiang MENG ; Liu ZHANG ; Faming TIAN ; Dacheng HAN ; Jie ZHENG ; Jun CAI
Chinese Journal of Tissue Engineering Research 2010;14(11):2081-2085
BACKGROUND:Simvastatin enhanced the expression of bone morphogenetic protein-2(BMP-2),which plays an anabolic role in bone metabolism and osteoblastic lineage differentiation.However,little is known about the molecular mechanism of simvastatin on regulation of bone marrow stromal cells differentiation.OBJECTIVE:To investigated the effect of simvastatin on osteoblastic differentiation of bone marrow stromal cells based on genetics level.METHODS:Bone marrow stromal cells derived from femur and tibia were cultured in different mediums with simvastatin or Vehicle for 7 days Following extraction and purification,mRNA was reverse-transcripted into cDNA.Fluorescence labelina was employed and the samples were then hybridized with oligonucleotide chip to screen the different genes,which were utillzed to analyze osteogenesis-related factors.Alkaline phosphatase and Von Kossa staining were performed at days 14 and 21,respectively.RESULTS AND CONCLUSIONS:At day 14,alkaline phosphatase-positive cells were more in the experimental group than control group.Von Kossa staining demonstrated that simvastatin could promote BMSCs osteoblastic differentiation and mineralization.Comparative analysis showed that 103 genes out of 22 575 rat genes had differential expression (≥2 fold or≤ 0.5 fold),and some genes were related to cell proliferation and ostoeblastic differentiation,including C/EBP δ,Cited,Ascl2,Ptpnl6,Wisp2,Tieg,etc.Simvastatin could induce osteoblastic differentiation of bone marrow stromal cells,involving in many osteogenetic-related genes.
4.Effect of simvastatin on bone formation and osteogenic differentiation of bone marrow stromal cells in young rats
Xiaoning LIU ; Liu ZHANG ; Faming TIAN ; Hui ZHANG ; Dacheng HAN ; Junqiang NIU ; Lei ZHANG
Chinese Journal of Tissue Engineering Research 2010;14(7):1152-1156
BACKGROUND: Recently simvastatin has been shown to stimulate osteogenic differentiation and bone formation, but there is no report about the effect of simvastatin on the bone development of young rats.OBJECTIVE: To evaluate the effects of simvastatin on osteogenic relative genes of proximal tibia trabecular bone and osteogenic differentiation of bone marrow stromal cells (BMSCs).METHODS: Twenty 1-week-old Spragua-Dawley young rats were randomly and equally divided into simvastatin and control groups. Rats in the simvastatin group were treated with a subcutaneous injection of simvastatin[5 mg/(kg·d)] for 2 weeks, while rats in the control group were treated with placebo for 2 weeks. The expressions of bone morphogenetic protein-2 (BMP-2), matrix metalloproteinase-13 (MMP-13), and vascular endothelial growth factor (VEGF) of trabecular bone in the tibia were analyzed by mmunohistochemicel staining. BMSCs harvested from the rat femur were osteogenic-differentiation cultured. Alkaline phosphatase (ALP) staining was performed on day 14, real-time PCR analysis was applied to investigate the BMP2, RUNX2,Osterix, MSX2, DLX3, DLX5 mRNA expressions during osteogenic differentiation in vitro on day 21, and von Kossa staining was detected on day 28.RESULTS AND CONCLUSION: ① There was no significant difference in the expressions of BMP-2, MMP-13, and VEGF between simvastatin and control groups. ② The percentages of ALP positive-stained cells were about 30% and there was no significant difference between the two groups (P > 0.05). ③There was no significant difference in the expressions of BMP-2,RUNX2, Osterix, MSX2, DLX3, DLX5 mRNA in osteoganic differentiation-induced BMSCs. ④ von Kossa staining demonstrated that dark brown calcified spots in various sizes were observed, but there was no significant difference in size and density between simvastatin and control groups. A subcutaneous injection of simvastatin[5 mg/(kg·d)] for 2 weeks could not remarkably affect osteogenic relative genes of bone trabecula and osteogenic differentiation of BMSCs.
5.Effect of preoperative thyroid dysfunction on the 30-day postoperative mortality and complications in elderly patients with hip fracture
Anhua LONG ; Zongyan XIE ; Dacheng HAN ; Jialong WANG ; Feifei ZHAO ; Lu JIN ; Xuefei WANG ; Yakui ZHANG
Chinese Journal of Orthopaedic Trauma 2021;23(3):216-220
Objective:To investigate the influence of preoperative thyroid dysfunction on the 30-day postoperative mortality and complications in elderly patients with hip fracture.Methods:A retrospective analysis was conducted of the 349 elderly patients with hip fracture who had been admitted to Department of Orthopedic Trauma, Beijing Luhe Hospital Affiliated to Capital Medical University from January 2018 to December 2019. They were 108 males and 241 females, with an average age of 76.3 years (from 60 to 104 years). There were 190 femoral intertrochanteric fractures and 159 femoral neck fractures. By the preoperative level of thyroid function, the patients were divided into a normal function group of 290 cases and a dysfunction group of 59 cases. The 2 groups were compared in terms of hospital stay, mortality and incidence of complications within 30 days postoperation.Results:In this cohort, the rate of 30-day postoperative mortality was 3.4%(12/349) and the incidence of 30-day postoperative complications 14.6%(51/349). The 2 groups were comparable because there was no significant difference between them in the preoperative general data except for the preoperative comorbidity of coronary heart disease ( P>0.05). In the dysfunction group, the hospital stay averaged (10.2±6.9) d, the rate of 30-d postoperative mortality 1.7%(1/59) and the incidence of 30-day postoperative complications 16.9%(10/59), which were insignificantly different from those in the normal function group [(10.7±7.5) d, 3.8%(11/290) and 14.1%(41/290), respectively] ( P> 0.05). Conclusion:Since preoperative thyroid dysfunction does not affect the 30-day postoperative mortality and postoperative complications in the elderly patients with hip fracture but no definite thyroid disease, routine thyroid function screening is not recommended for them.
6.Comparative study of open surgery and arthroscopic assisted surgery in management of knee dislocations
Jinwen HE ; Dacheng ZHAO ; Bin GENG ; Yuanjun TENG ; Chengjun ZHANG ; Meng WU ; Xiangdong YUN ; Hua HAN ; Yayi XIA
Chinese Journal of Trauma 2021;37(2):114-121
Objective:To compare the efficacy of open surgery and arthroscopic assisted surgery in treatment of knee dislocations.Methods:A retrospective case-control study was conducted to analyze the clinical data of 80 patients with knee dislocations admitted to Second Hospital of Lanzhou University from May 2013 to September 2019, including 59 males and 21 females, aged 18-66 years [(42.5±11.6)years]. Open multiple ligament reconstruction was performed in 49 patients (open group) and arthroscopic assisted multiple ligament reconstruction was performed in 31 patients (arthroscopic group). The postoperative hospitalization days, incidence of complications, time needed for recovery of knee range of motion (>0°, >90°, >120°), and time to complete weight-bearing were compared between the two groups. The Lysholm score, international knee documentation committee (IKDC) subjective knee form, Tegner activity level, score of the MOS item short-form health survey (SF-36), patient satisfaction and knee range of motion were compared between the two groups at the last follow-up.Results:All the patients were followed up for 1.2-7.4 years [(3.8±1.5)years]. There was no significant difference in postoperative hospitalization days or incidence of complications between the two groups ( P>0.05). No significant difference was found in time needed for recovery of knee range of motion (>0°, >120°) or time to complete weight-bearing ( P>0.05). The time needed for recovery of knee range of motion (>90°) was 90(60, 90)days in open group and 60(30, 90)days in arthroscopic group ( P<0.05). At the last follow-up, there was no significant difference in Lysholm score, IKDC subjective score, Tegner activity level, SF-36 score, or patient satisfaction between the two groups ( P>0.05). At the last follow-up, the knee range of motion was 120°(90°, 130°) in open group and 135°(120°, 140°) in arthroscopic group ( P<0.05). Conclusion:For treatment of knee dislocations, open surgery and arthroscopic assisted surgery have similar results in the long-term effect, while arthroscopic assisted surgery has benefits in early rehabilitation and ultimately better knee range of motion.
7.Robot-assisted thoracic surgery versus video-assisted thoracic surgery for early-stage lung cancer: A case control study
MA Jilong ; JIN Dacheng ; HAN Songchen ; CHEN Meng ; GOU Yunjiu
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2019;26(1):48-52
Objective To evaluate the curative effect of robot-assisted thoracic surgery (RATS) and video-assisted thoracic surgery (VATS) for early stage lung cancer patients. Methods We retrospectively analyzed the clinical data of 80 patients with radical resection of lung cancer in Gansu Provincial Hospital between January 2016 and December 2017. The patients were divided into two groups: 43 patients in a VATS group and 37 in a RATS group. There were 51 males and 29 females. Data were processed with STATA v14.0 statistical software. Results There were significant differences in the operative time, duration of hospital stay and operative costs between the two groups. In the RATS group, operative duration was longer (172.21 minutes versus 162.20 minutes, P=0.018), cost was higher (74 076.69 yuan versus 54 814.73 yuan, P<0.001), and required significantly shorter hospital stay (8.27 daysversus 10.76 days, P=0.001) compared with those of the VATS group. There was no statistical difference between the two groups in terms of conversion (3 versus 0, P=0.144), blood loss during operation (61.29 ml versus 90.63 ml, P=0.213), dissected lymph node number (17.38 versus 12.63, P=0.095), drainage volume (1 406.76 ml versus 1 514.60 ml, P=0.617) and the drainage time (7.92 days versus 7.20 days, P=0.440). Conclusion In the early lung cancer patients who underwent thoracic surgery, the postoperative hospitalization time of the RATS group is shorter than that of the VATS group, and the operation time is longer than that of the VATS group. The other short-term surgical indexes are similar to those of thoracoscopic surgery. However, the robot has great advantages in the treatment of patients with difficult lymph node dissection, serious pleural adhesion and complicated anatomical relationship.
8.A comparison of clinical effects of removing foreign bodies from esophagus by rigid esophagoscope and flexible esophagoscope
GOU Yunjiu ; MA Jilong ; HAN Songchen ; JIN Dacheng ; CHEN Meng ; WANG Bing ; BAI Qizhou
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2019;26(12):1180-1184
Objective To collect the data of esophageal foreign body patients, and to evaluate the clinical effects of two different surgical methods in our hospital. Methods The clinical data of 294 patients who were treated in Gansu Provincal Hospital from January 2012 to June 2018 were analyzed retrospectively. The clinical data were collected and analyzed by SPSS 22.0. In order to to evaluate the efficacy of flexible esophagoscope (FE) and rigid esophagoscope (RE) in the treatment of esophageal foreign bodies.The patients were divided into two groups: a RE group including 118 patients with 62 males and 56 females at age of 6 (3-37) years and a FE group including 176 patients with 84 males and 92 females at age of 6 (3-59) years. Results There was no significant difference in age, age stratification, gender and foreign body type between the two groups. There was a statistical difference in the initial clinical symptoms (P=0.041) or in esophageal foreign bodies position (P=0.037) between the two groups. The success rate of foreign body removal was similar between the two groups (P=0.632). The success rate was 88.9% (105/118) in the RE group, 87.5% (154/176) in the FE group. The operation time was significantly longer in the RE group than that in the FE group (10.8 ±17.4 min vs. 17.5±21.6 min, P<0.001). The postoperative hospitalization time in the RE groups was longer than that in the FE group (21.5 ±24.2 hours vs. 12.5 ±21.3 hours, P<0.05). There was a statistical difference in the incidence of postoperative complications between the two groups (P=0.034). In the RE group, the main complication was mucosal edema (15.3%). And the rate of bleeding was higher (15.9%) in the FE group. There were 30 patients (25.5%) in the RE group with minor postoperative complications versus the FE group with 40 patients (22.7%); and 1 patient (0.8%) in the RE group with severe complications versus the FE group with 5 paients (2.8%). Conclusion Based on the analysis of this study, it is found that RE has higher safety. But the indications are strict, the professional requirements of the operator and the selection of patients are stronger. The FE is convenient to use, the operation crowd is wide, and the suitable crowd is wide. Therefore, for specific patients, after improving the relevant examination and preoperative evaluation of patients, clinicians need to choose appropriate surgical methods to ensure the success of the operation, and reduce the postoperative complications as far as possible.
9.Perioperative efficacy and safety of da Vinci robot-assisted bronchial sleeve lobectomy
Siyuan ZHANG ; Xinchun DONG ; Yunjiu GOU ; Songchen HAN ; Meng CHEN ; Dacheng JIN
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2020;27(10):1145-1149
Objective Through the perioperative outcome analysis of da Vinci robot-assisted sleeve lobectomy, to clarify its efficacy and safety. Methods A retrospective analysis was performed on 10 patients with centrally located lung cancer undergoing robot-assisted sleeve lobectomy from March to December 2019 in our center, including 9 males and 1 female, aged 45-67 (55.0±8.9) years. Preoperative imaging and bronchoscopy showed central non-small cell lung cancer, involving the right upper lung in 3 patients, right lower lung in 2 patients, the left upper lung in 4 patients, and left lower lung in 1 patient. The operation time, Docking time, intraoperative blood loss volume, bronchial anastomosis time, number of dissected lymph nodes, drainage volume and postoperative hospital stay were analyzed. Results The da Vinci robot-assisted bronchial sleeve lobectomy was completed smoothly on 10 patients. The operation time was 135-183 (157.8±14.3) min, Docking time 6-15 (10.0±2.9) min, intraoperative blood loss volume 55-250 (124.5±61.8) mL, bronchial anastomosis time 17-40 (27.7±7.3) min, the number of dissected lymph nodes 16-23 (19.7±2.8), the drainage volume 200-600 (348.0±148.4) mL and postoperative hospital stay 7-11 (8.7±1.6) d. All patients had no bronchopleural fistula, pulmonary infection or atelectasis, and there was no perioperative death. Postoperative pathological findings were all squamous cell carcinoma. Conclusion Da Vinci robot-assisted sleeve lobectomy is safe and effective.
10.Advances and prospects in the application of robotic surgery system in the treatment of esophageal cancer
WANG Bing ; BAI Qizhou ; JIN Dacheng ; MA Jilong ; HAN Songchen ; CHEN Meng ; GOU Yunjiu ; WANG Shumin
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2019;26(7):715-722
At present, the application of the robot assisted surgery system in the surgical treatment of esophageal cancer is gradually emerging, and it is more and more widely used and recognized in the field of surgery. According to the domestic and foreign literatures, the robot has many advantages, and robotic assisted esophageal cancer surgery has been proved to be safe and effective, and its short-term efficacy is significantly better than thoracotomy. Other studies have shown that in long-term follow-up, the effect is comparable to video-assisted thoracoscopic surgery. In this paper, the author are systematically reviewed the development history of the robot assisted surgery system, the effect of robotic assisted esophagectomy on safety, surgical method, short-term efficacy and long-term prognosis. The traditional open surgery and thoracoscopic laparoscopic esophagectomy has been carried on the detailed comparison to provide some advice and theoretical basis for esophageal cancer surgery robot system.