Objective To explore the proper dosage of establishment of stable hepatic oval cells （HOC） prolif-eration model by using 2-acetaminofluorene （2-AAF） combined with two-third partial hepatectomy （2/3 PH） surgery， and to explore isolated and cultured method of HOC in vitro. Methods The 174 Wistar rats were randomly divided into 4 experimental groups （each group enrolled 30 rats）， saline group （n=30）， and untreated group （n=24）. Rats of 4 experi-mental groups were underwent gavage of 5， 10， 15， and 20 mg/（kg ? d） 2-AAF， corresponding to the groups from No.1 to No.4 group. Rats of saline group received saline gavage and rats of untreated group didn’t received any treatment. A standard 2/3 PH surgery was performed on the 5th day after gavage， then the same gavage method was still administrated as preoperation untill rats were sacrificed. The liver tissues of 6 selected rats were adopted and identified by HE staining and immunohistochemical staining on 4， 8， 12， and 16 days after PH for observation of the proliferation of HOC in every group， on 4 days， levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） were tested in addition. HOC were isolated and purified by collagenase perfusion method and percoll gradient centrifugation. Results The surv-ival rates of untreated group，saline group，No.1 group，No.2 group，No.3 group，and No.4 group were 100% （24/24），93% （28/30），93% （28/30），90% （27/30），90% （27/30），and 80% （24/30） respectively. Compared with the saline group and untreated group， the levels of serum ALT and AST increased significantly in No.2， No.3， and No.4 group on the 4th day after PH （P＜0.05）. The results of HE staining showed that No.2， No.3， and No.4 group were observed visibly different level of damage at liver tissue， and the proliferation level of HOC were most obviously in No.3 and No.4 group. The results of immunohistochemical staining revealed that proliferation cells were positively expressed oval cell marker-6 （OV-6）. The number of OV-6 positive cells were increased significantly with the increase of dosage of 2-AAF between 4 days and 12 days after operation， and proliferation levels were related with dosages of 2-AAF （P＜0.05）. In all cultured cells， 80% of cells were OV-6 positive cells after isolation and culture by using collagenase perfusion method and percoll gradient centrifugation. Conclusions The methods of gavage of 2-AAF at 15 mg/（kg ? d） combined with 2/3 PH surgery can establish the HOC proliferation model on the 12th day， as well as the rats have lower mortality and better tolerance， especially. The collagenase perfusion method and percoll gradient centrifugation can be used to isolate HOC effectively.
Objective To evaluate the clinical experience and skills of laparoscopic totally extraperitoneal hernia repair with non-stapling of mesh and without using a balloon dissection in order to spread and popularize this kind of operation. Methods Under general anesthesia, 32 patients (42 sides) with inguinal hernias were repaired by laparoscopic totally extraperitoneal hernia repair with non-stapling of mesh and without using a dissection balloon from August 2005 to December 2007. Results All of operations were successfully performed. The operative time was 60-120 minutes, the blood loss was 20-60 ml, and the length of postoperative hospitalization was 3 days. All cases were followed up for 8 to 18 months and found no recurrence. Conclusion Laparoscopic totally extraperitoneal hernia repair with non-stapling of mesh and without using a balloon dissection is feasible, reliable and effective, offering a low recurrence rate, while its price is higher than tension-free herniorrhaphy by traditional method.
【Abstract】 Objective To explore good methods for isolation and purification of rat islets. Methods The isletswere isolated from male SD rat pancreata by a collagenase perfusion method and purified by a modified method: added 4 kinds of Euro-Ficoll of different densities (F1: D=1.132, F2: D=1.108, F4: D=1.069, F5: D=1.023), discontinuous density gradient centrifuge the tube at 2 000 r/min for 20 minutes at 4℃ , then the islets between F1 and F2 were collected. The purity of islets was assessed by dithizone staining with islets counted and scored for size. Islets viabil ity was assessed by fluorescin diacetate / propidium iodide. The function of purified islets was judged by the test of insul in release and islets transplantation. Results After an improved method for optimized isolation and purification, (920±122) IEQ purified islets were obtained from one rat. Both the purity and viabil ity of islets were over 90%. The amount of insul in secretion was (18.25±0.32) mU/L and (36.70±3.57) mU/Lat 2.2 mmol/ L and 22.2 mmol/L concentration of glucose respectively, there was significant difference between the two phases(P lt; 0.05). The insul in release index was 2.01±0.15. Under 1 000 IEQ islets transplantation, the normal glucose level could beremained in diabetic rats. Conclusion High purity and high viabil ity islet cells can be got through improved collagenase perfusion and centrifugation on gradients method.
ObjectiveTo understand the latest progress of enrichment technology of circulating tumor cells (CTCs), and summarize the principle, advantages and disadvantages of various enrichment technologies and their applications in primary liver cancer (PLC). MethodThe literature relevant to the enrichment methods of CTCs in the PLC was reviewed and summarized. ResultsThe clinical significances of CTCs in the early diagnosis and staging, hierarchical diagnosis and treatment, and efficacy monitoring of patients with PLC had been recognized. There were many separation and enrichment technologies for CTC, which were mainly based on the differences of physical and biochemical characteristics, as well as the combination of enrichment methods with various principles. Each enrichment method had corresponding advantages and disadvantages, and few enrichment methods for CTC was applied to PLC. ConclusionsAlthough many problems need to be solved in enrichment method of CTCs at present, it is believed that the existing problems will be solved one by one with continuous improvement of technology. And CTC detection is expected to apply in clinical, so as to provide more efficient diagnosis and treatment methods for patients with PLC.
Kidney transplantation is an ideal treatment for patients with end-stage renal disease. Circulating alloantibodies against donor human leukocyte antigens and blood group antigens can impair allografts, shorten allograft survival, and limit access to kidney transplantation. Furthermore, the presence of donor specific antibodies is associated with increased incidence of antibody-mediated rejection and decreased graft survival following transplantation. Plasmapheresis, an extracorporeal therapy directed at removing plasma proteins that has been found to minimize the effects of perioperative sensitization in kidney transplantation. Plasmapheresis enables transplantation across the barrier of ABO blood group incompatibility. In addition, it is also an important approach for the treatment of antibody-mediated rejection. Therefore, studying the application of plasmapheresis in perioperative period of kidney transplantation is expected to increase the chance of transplantation and improve the outcomes following transplantation. This article introduces the application of plasmapheresis in the perioperative period of kidney transplantation.
ObjectiveTo summarize the progress in treatment of pubic symphysis diastasis. MethodsRelated literature concerning treatment of pubic symphysis diastasis was extensively reviewed and comprehensively analyzed in terms of anatomy, biomechanics, and treatment. ResultsThere are many fixation methods for treatment of pubic symphysis diastasis, which aims at restoring the stability of the anterior pelvic ring. External fixator is often used as a temporary fixation; tension band wire has been abandoned due to its poor biomechanical stability; screw loosening and plate breakage often appears when a single reconstruction plate is used; box plate significantly increases the biomechanical stability of anterior pelvic ring but it leads to a considerable surgical trauma; locking plate has been used for pubic symphysis diastasis recently, especially for osteoporotic fractures; percutaneous cannulated screw has the advantages of less trauma, less bleeding, and good stability, so it is good choice for treatment of pubic symphysis diastasis. ConclusionThere is no uniform standards about the treatment of pubic symphysis diastasis, but the minimally invasive treatment is an undeniable trend. Percutaneous cannulated screw has achieved satisfactory effectiveness, however, its biomechanical stability and anatomic channels need to be further studied.
ObjectiveTo evaluate the effects of sevoflurane and propofol on preoperative implicit and explicit memories in general anaesthesia patients of elective surgery. MethodsThe surgical inpatients in Sichuan Provincial People's Hospital were enrolled from December 2013 to May 2014, and were randomly divided into three groups (S, P, M). In Group S, anesthesia was induced and maintained with sevoflurane. In Group P, anesthesia was induced and maintained with propofol. Midazolam was not utilized throughout the whole anaesthesia for the above groups. Patients in Group S and Group P were given a list of test materials to remember and listen before the anesthesia. Within 12 to 36 hours after operation, memory was assessed, based on the Buchner's model applied on the process dissociation procedure (PDP) using a phrases task. The Group M was given the same test materials, and received test with the PDP in 12 to 36 hours before surgery. Value A and value R were used to represent the implicit memory score and the explicit memory score, respectively. ResultsA total of 150 patients were included, and 50 cases were included in each group. During testing, 2 cases were excluded, 3 cases were loss to follow-up, so finally 49 cases were included in the Group S, 47 cases in the Group P and 49 cases in the Group M. The results showed that there were significant differences in the implicit memory score (A) and the explicit memory score (R) among the three groups (all P values <0.05). The explicit memory score (R) of the Group M was higher than those of the Group P and Group S (all P values <0.05), the implicit memory score (A) in the Group M was higher than those of the Group S and Group P (all P values <0.05), and the implicit memory score (A) in the Group S was higher than that of the Group P (P<0.05). ConclusionPropofol and sevoflurane can decrease the score of explicit memory after anesthesia within 12 to 36 hours, and there are no significant differences in explicit memory between the two drugs. Both propofol and sevoflurane can decrease the score of implicit memory, but the influence of sevoflurane on the implicit memory is less than propofol within 12 to 36 hours.
【摘要】 目的 通过比较两种原代人脐静脉内皮细胞的分离培养方法并对细胞特异性抗原进行鉴定,探索提高原代内皮细胞体外培养存活率及纯化率的方法。 方法 采用一次性无菌注射器向人脐静脉灌注消化液,消化液的浓度和消化时间分别025%(质量体积比)胰蛋白酶,10 min和01%(质量体积比)胶原酶Ⅱ,15 min。通过在倒置显微镜下观察细胞的形态特点和用免疫荧光染色的方法对细胞进行鉴定,比较两种消化方法的优劣。 结果 01%胶原酶Ⅱ,15 min的消化方法较025%胰蛋白酶,10 min对原代人脐静脉内皮细胞有更好的分离效果,活细胞数量多且细胞纯度较高。免疫荧光染色结果表明细胞内有Ⅷ因子相关抗原表达。结论 胶原酶Ⅱ可以有效分离脐静脉内皮细胞,最佳消化条件是01%胶原酶Ⅱ,37℃,15 min。【Abstract】 Objective To explore the optimal method for primary culture of human umbilical vein endothelial cells (HUVECs). Methods HUVECs were prepared from human umbilical cords by 01% collagenase Ⅱ digestion for 15 minutes and 025 trypsinase digestion for 10 minutes,respectively. HUVECs were observed under inverted microscope and identified by immunofluorescence.The two methods of digestion were compared. Results More HUVECs were harvested through the method of 01% collagenase Ⅱ for 15 minutes,which expressed Ⅷ related antigen. Conclusion The method of 0.1% collagenase Ⅱ digestion for 15 minutes is a better choice to isolate HUVECs.
The impulsive electroencephalograph (EEG) noises in evoked potential (EP) signals is very strong, usually with a heavy tail and infinite variance characteristics like the acceleration noise impact, hypoxia and etc., as shown in other special tests. The noises can be described by α stable distribution model. In this paper, Wigner-Ville distribution (WVD) and pseudo Wigner-Ville distribution (PWVD) time-frequency distribution based on the fractional lower order moment are presented to be improved. We got fractional lower order WVD (FLO-WVD) and fractional lower order PWVD (FLO-PWVD) time-frequency distribution which could be suitable for α stable distribution process. We also proposed the fractional lower order spatial time-frequency distribution matrix (FLO-STFM) concept. Therefore, combining with time-frequency underdetermined blind source separation (TF-UBSS), we proposed a new fractional lower order spatial time-frequency underdetermined blind source separation (FLO-TF-UBSS) which can work in α stable distribution environment. We used the FLO-TF-UBSS algorithm to extract EPs. Simulations showed that the proposed method could effectively extract EPs in EEG noises, and the separated EPs and EEG signals based on FLO-TF-UBSS were almost the same as the original signal, but blind separation based on TF-UBSS had certain deviation. The correlation coefficient of the FLO-TF-UBSS algorithm was higher than the TF-UBSS algorithm when generalized signal-to-noise ratio (GSNR) changed from 10 dB to 30 dB and α varied from 1.06 to 1.94, and was approximately equal to 1. Hence, the proposed FLO-TF-UBSS method might be better than the TF-UBSS algorithm based on second order for extracting EP signal under an EEG noise environment.
Objective To investigate the clinical efficacy between suture button plate and cortical screw in the treatment of distal tibiofibular syndesmosis seperation. Methods The clinical data and follow-up data 1 week and 2, 4, 6 weeks and 3, 6, 12 months after operation of patients with distal tibiofibular syndesmosis seperation combined with or without ankle fracture who were admitted to hospital from March 2011 to May 2014 were retrospectively collected. According to the fixation ways of distal tibiofibular syndesmosis, the patients were divided into the suture button plate group and the cortical screw group. The operation time, amount of blood loss, combined ankle fracture healing time, American Orthopaedic Foot and Ankle Society (AOFAS) scores at the last follow-up and complications were compared between the two groups. Results A total of 52 patients were enrolled, including 28 cases of suture button plate group and 24 cases of cortical screw group. There were no significant difference in operation time or amount of blood loss between the two groups (P>0.05). All patients were followed up for 12–30 months averaged 14.5 months. All incisions healed by the first intention. The combined ankle fracture healing time in the suture button plate group was (11.2±2.0) weeks, while that was (11.0±2.1) weeks in the cortical screw group, with no significant difference between the two groups (P>0.05). In the suture button plate group, there was no loosening or rupture of internal fixation, while distal tibiofibular syndesmosis seperation was found again in one case 3 weeks after operation, and cured by reoperation at last. In the cortical screw group, rupture of screws was found in 2 cases, which occurred 9 and 11 weeks after operation respectively, and the broken screws were taken out when the fractures were healed and the internal fixation was removed; distal fibiofibular syndesmosis seperation was found again in one case 6 weeks after operation, and no treatment was given due to old age and low demand. At the last follow-up, the AOFAS score in the suture button plate group was 84.6±10.2, while the score in the cortical screw group was 83.7±12.5, with no significant difference between the two groups (P>0.05). Conclusions Suture button plate and cortical screw can effectively treat distal fibiofibular syndesmosis separation. The risk of suture button plate loosening or rupture is poor. Suture button plate fixation can meet the need of early functional exercises, and avoid the routine removal by reoperation.