[Objective] To explore the temperature parameters affecting the polybrene test and determine the optimal temperature conditions for detecting unexpected antibodies of the Rh system. [Methods] The reaction of IgG human anti-D antibody with different dilutions (undiluted, 1∶2, 1∶4, 1∶8, 1∶16, 1∶32，1∶64) with D antigen-positive red blood cells was detected by manual polybrene test (MPT). Different temperatures (25℃ and 37℃) were set, and the reaction time with low ionic medium was 4 minutes. The agglutination integral value of anti-D and red cell depolymerization time were compared to observe the effect of enhanced agglutination reaction, thereby establishing the test temperature reaction conditions for enhancing the MPT. The same reaction condition was applied to 36 blood samples containing unexpected antibodies of the Rh system, and the effect of enhanced MPT was observed in comparison with the polybrene method and the antiglobulin test (column agglutination). [Results] With all other conditions held constant, when low ionic medium was added, the incubation temperature of 25℃ and 37℃ resulted in different total agglutination integral values for anti-D (20.9±2.025 vs 25.5±2.635), and the comparison showed a significant difference (P<0.05). When the antibody dilution was 1∶16, the incubation temperature of 25℃ and 37℃ resulted in different agglutination integral values (3.9±0.738 vs 5.8±0.632), and the comparison showed a significant difference (P<0.05). Erythrocyte depolymerization time (62.8±8.149 vs 90.1±10.713) was significantly different (P<0.05). At a dilution of 1∶32, the incubation temperatures of 25℃ and 37℃ resulted in different agglutination integral values (2.5±0.527 vs 4.3±0.675), as well as different red blood cell dissociation times (35.4±7.792 vs 57.4±10.885)(P<0.05), and the comparison showed a significant difference (P<0.05), with no differences observed in the other groups. In the detection of 36 Rh system unexpected antibody samples, when the antibody titer was ≤2, the enhanced polybrene method had a higher positive rate, and when the antibody titer was ≥4, the detection rates of the three methods were consistent. [Conclusion] The reference temperature condition for the modified MPT is incubation at 37℃ for 4 min after the addition of low ionic medium. The application of this temperature condition to unexpected antibody samples of Rh system could achieve a significant enhancement effect, thereby increasing transfusion safety for the treatment of emergency patients, and is worth popularizing.

Objective:To evaluate the application effect of management mode based on constraint theory for medical equipment in equipment management of vascular surgery. Methods:Based on five-step method of constraint theory,an operation evaluation system was formulated. An information management mode of medical equipment was constructed to manage quality of medical equipment in department of vascular surgery. A total of 60 sets used medical equipment in the Department of Vascular Surgery at The Second Affiliated Hospital of Xi'an Jiaotong University from 2021 to 2022 were selected. The conventional management mode was adopted to manage these equipment between January to December 2021,while equipment was managed by using medical equipment management mode based on constraint theory (constraint theory management) of the department of vascular surgery from January to December 2022. The quality scores of operation and maintenance of equipment,average incidence rates of risk,configuration effectiveness,and timeliness of equipment management were compared between the two management modes. Results:After constraint theory management was implemented,the average scores of operation and maintenance of equipment,which included success starting-up,operation,routine maintenance and fault repair,were respectively (90.69±8.65),(89.65±4.65),(88.65±5.63),and (90.58±6.01),all of which were higher than those of conventional management mode. The differences of them between two modes were statistically significant (t=13.482,17.867,14.552,14.449,P<0.05). The average incidence rates of the risks of testing equipment,treatment,monitoring equipment and auxiliary equipment of adopting constraint theory mode management were (0.89±0.22)％,(0.84±0.23)％,(0.74±0.32)％ and (0.75±0.32％),all which were lower than those of the conventional management mode,with statistically significant differences (t=9.162,17.745,19.469,15.019,P<0.05). The average utilization rate of equipment and the average amplification of cost benefit of equipment were respectively (88.69±5.54)％ and (50.36±2.69)％ after constraint theory mode was adopted,which were significantly higher than those of conventional management mode,and the average floating rate of using equipment of constraint theory mode was (1.23±0.97)％,which was lower than that of conventional management model,the differences of them between two modes were statistically significant (t=14.174,21.564,11.811,P<0.05). The average times of emergency management for risk events included the safety of power consumption of equipment,safety of diagnosis and treatment,biological safety and radiation safety were respectively (2.36±0.64)h,(3.26±0.78)h,(4.23±1.24)h and (2.26±1.23)h after the constraint theory mode was adopted,all of which were significantly less than those of conventional management mode,and the differences of them between two modes were statistically significant (t=16.704,13.421,9.029,13.971,P<0.05). Conclusion:The application of the management mode based on the constraint theory of medical equipment in department of vascular surgery can improve the quality of operation and maintenance of equipment for safety,and shorten the emergency response time of equipment,and reduce the incidence of equipment risk,and strengthen the effect of configuration for equipment.

Objective:To evaluate the application effect of management mode based on constraint theory for medical equipment in equipment management of vascular surgery. Methods:Based on five-step method of constraint theory,an operation evaluation system was formulated. An information management mode of medical equipment was constructed to manage quality of medical equipment in department of vascular surgery. A total of 60 sets used medical equipment in the Department of Vascular Surgery at The Second Affiliated Hospital of Xi'an Jiaotong University from 2021 to 2022 were selected. The conventional management mode was adopted to manage these equipment between January to December 2021,while equipment was managed by using medical equipment management mode based on constraint theory (constraint theory management) of the department of vascular surgery from January to December 2022. The quality scores of operation and maintenance of equipment,average incidence rates of risk,configuration effectiveness,and timeliness of equipment management were compared between the two management modes. Results:After constraint theory management was implemented,the average scores of operation and maintenance of equipment,which included success starting-up,operation,routine maintenance and fault repair,were respectively (90.69±8.65),(89.65±4.65),(88.65±5.63),and (90.58±6.01),all of which were higher than those of conventional management mode. The differences of them between two modes were statistically significant (t=13.482,17.867,14.552,14.449,P<0.05). The average incidence rates of the risks of testing equipment,treatment,monitoring equipment and auxiliary equipment of adopting constraint theory mode management were (0.89±0.22)％,(0.84±0.23)％,(0.74±0.32)％ and (0.75±0.32％),all which were lower than those of the conventional management mode,with statistically significant differences (t=9.162,17.745,19.469,15.019,P<0.05). The average utilization rate of equipment and the average amplification of cost benefit of equipment were respectively (88.69±5.54)％ and (50.36±2.69)％ after constraint theory mode was adopted,which were significantly higher than those of conventional management mode,and the average floating rate of using equipment of constraint theory mode was (1.23±0.97)％,which was lower than that of conventional management model,the differences of them between two modes were statistically significant (t=14.174,21.564,11.811,P<0.05). The average times of emergency management for risk events included the safety of power consumption of equipment,safety of diagnosis and treatment,biological safety and radiation safety were respectively (2.36±0.64)h,(3.26±0.78)h,(4.23±1.24)h and (2.26±1.23)h after the constraint theory mode was adopted,all of which were significantly less than those of conventional management mode,and the differences of them between two modes were statistically significant (t=16.704,13.421,9.029,13.971,P<0.05). Conclusion:The application of the management mode based on the constraint theory of medical equipment in department of vascular surgery can improve the quality of operation and maintenance of equipment for safety,and shorten the emergency response time of equipment,and reduce the incidence of equipment risk,and strengthen the effect of configuration for equipment.

Objective To construct engineered corneal epithelium from embryonic stem cells (ESCs) using Rock inhibitor combined with hypoxia-normoxia culture condition. Methods Human ESC line H1 was induced to differentiate into epithelial-like cells by addition of retinoic acid (RA) and bone morphogenetic protein 4(BMP4) in the differentiation medium under the adherent culture condition. The ESCs derived epithelial-like cells were expanded in the mixed medium of SHEM and KSFM with the mixture ratio of 1 : 2 with or without Rock inhibitor Y27632. The H1 derived epithelial-like cells were seeded on the denuded ammonic membrane to construct engineered corneal epithelium under hypoxia,normoxia and hypoxia-normoxia culture conditions,respectively. The inducted effect of ESCs into epithelial-like cells,the expansion ability of the epithelial-like cells and the characteristics of the constructed engineered corneal epithelium were evaluated by morphological observation, real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and immunofluorescence technology. Results Compared with the control group,the relative expressions of ESCs marker Oct4 mRNA, Notch signaling pathway related factors Notch1 and Jagged1 mRNA,and Wnt signaling pathways related factors c-myc and Cyclin D1 mRNA were significantly reduced, and the relative expressions of cutaneous ectoderm markers p63 and K18 mRNA were significantly increased at day 8 after induction in the induced group,with significant differences between them (t =14.63,20.15,93.50,11.60, 19.30,18.44,22.63;all at P<0.05). Compared with the without Y27632 group,the relative expressions of p63 and K14 mRNA,Notch signal pathway receptor Notch1 and Jagged1 mRNA were significantly increased,and Wnt signaling pathways downstream targeted gene c-myc and CylinD1 mRNA were significantly decreased at day 8 after induction in the Y27632 group,with significant differences between them (t =20.29,59.22,2.90,39.59,5.32,10.14;all at P<0.05),and the relative expression of K18 mRNA in the two groups was not significantly changed(t=1.38,P>0.05). The ESCs derived epithelium and constructed under hypoxia-normoxia culture condition showed more obvious stratification and tighter cell arrangement in comparison with those cells cultured in consistent hypoxia culture condition or normoxia culture condition. Epithelial markers Pan-CK and K18 as well as epithelial progenitor cell markers p63 and K14 expressed in the whole cell layers of the ESCs derived epithelium constructed under hypoxia-normoxia culture condition. Conclusions The addition of Y27632 enhances the proliferation ability of H1 derived epithelial cells and actives Notch signaling pathway and inhibits Wnt signaling pathway. The culture and construction in the expansion medium with Y27632 under the hypoxia-normoxia culture condition can promote the stratification of H1 derived engineered corneal epithelium.

Objective To clarify the role of nuclear factor κB(NF-κB) signaling pathway in pathogenesis of Kashin-Beck disease(KBD) by observing the expression of NF-κB p65 in the whole blood samples of patients with KBD and controls,and the expression of NF-κB p65 in C28/I2 chondrocyte, and to analyze the role of NF-κB p65 molecule in chondrocyte apoptosis. Methods Through a case-control study, 161 patients with KBD (KBD group) were selected from Xunyi, Yongshou, Changwu, Linyou, Qianyang and Long counties in KBD endemic areas and 312 healthy people(control group) were matched by age and sex in Shaanxi Province. Venous blood samples were collected from patients and healthy controls, which were anticoagulated and used for determination of NF-κB p65 protein.According to the group design,the model of C28/I2 chondrocyte oxidative damage was established.The experiments were divided into 4 groups including control group(C), tBHP injury group (O, tBHP 300.00 μmol/L), low selenium pre-protection group (OS1, 0.05 mg/L Na2SeO3+ 300.00 μmol/L tBHP), and middle selenium pre-protection group(OS2, 0.10 mg/L Na2SeO3+ 300.00 μmol/L tBHP). Then, cell apoptosis was detected by Hoechst 33342 and reactive oxygen species (ROS) was detected by dichlorfluorescein(DCF) method. The protein was extracted by Trizol method, then protein expression level of NF-κB p65 molecule was detected by Western blotting in whole blood samples and C28/I2 chondrocyte. Results The differences in age and sex were not statistically significant between KBD group and control group (t = 0.336, P > 0.05; χ2= 0.407, P > 0.05). The protein expression level of NF-κB p65 in KBD group was 1.835 times as high as that of control group (KBD:0.167 ± 0.026, control: 0.091 ± 0.014, t = 5.147, P < 0.01). Under the fluorescence microscope, chondrocyte showed strong blue fluorescence in tBHP group and the level of ROS(1.219 ± 0.104) was higher than those of low and middle selenium pre-protection groups(0.832 ± 0.077, 0.635 ± 0.070, P < 0.05).The protein expression level of NF-κB p65 in tBHP group (1.563 ± 0.351) was higher than that of control group (0.451 ± 0.069, P < 0.05), and protein levels of NF-κB p65 had a decreasing tendency in low and middle selenium pre-protection groups compared to tBHP group. Conclusion The NF-κB signaling pathway is up-regulated in KBD patients, moreover, chondrocyte experiments show that cell apoptosis is mediated via upregulation of NF-κB p65,which suggests NF-κB signaling pathway may play an important role in pathogenesis of KBD.

Objective: To assess clinical and radiographic outcomes of short implants (length six mm) in the posterior region and early-loading with splinted fixed dental prostheses. Methods: Forty-five patients were recruited at 3 centers and 2-3 implants (diameter 4 mm, length 6 mm) were implanted in each case. Totally, ninty-five implants were placed, using one-stage surgery protocol and loaded with a screw-retained splinted fixed prosthesis 6 weeks later. Clinical and radiographic examinations were performed preoperatively, post-surgery, at loading, and 6, 12 and 24 months after prosthesis placement. Results: Four implants were removed before loading. A total of 36 patients with 76 implants completed the whole treatment and 2 year follow-up according to the research design. No implant was lost after loading, and the 2-year survival rate of the implant was 100%(76/76). In all cases, the response of swelling and pain after operation was mild, and the bone resorption around the implants was not obvious at 6 months [(0.04±0.29) mm], 1 year [(-0.11±0.44) mm], and 2 years [(0.00±0.53) mm] after loading. Bone loss less than 1.00 mm was found only in 26% (18/69) implants after 2 years of loading, and the height of the alveolar bone around 17% (12/69) of the implant increased. During 2-year follow-up, the probing depth of the implants increased by nearly 0.50 mm, but there was no clinical significance (P>0.05). Conclusions The 2-year results of this study showed that the 6-mm-long implants can be a predictable procedure, especially in situations with limited bone height in posterior edentulous regions.

Objective: To compare the peri-implant tissue stability between immediate implant and delayed implant in maxillary anterior region after loading 2 years.Methods: In the study, 38 patients with single anterior tooth loss in the Second Clinical Division of Peking University School and Hospital of Stomatology from October 2010 to December 2011 were enrolled , and 43 implants were inserted .The gin-gival contour was induced using implant-supported temporary crowns prior to restoration till permanent prostheses delivered .The gingival papilla height , labial gingival margin level and peri-implant bone level were measured immediately after the permanent restoration and 2 years later .Results: In the study , 16 patients were treated by immediate implant for 17 implants;22 patients were treated by delayed im-plant for 26 implants .The implant stability quotient ( ISQ ) value of the 2 groups showed no significant difference before permanent restoration (P>0.05).In all the cases after loading 2 years, the average mesial gingival papilla height in the implant area of the immediate group and delayed group increased by (0.15 ±0.42) mm and (0.06 ±0.65) mm, respectively;the distal gingival papilla height increased by (0.06 ±0.50) mm and (0.02 ±0.57) mm respectively;while the labial gingival margin level shrinka-ges were (0.15 ±0.23) mm and (0.15 ±0.46) mm, respectively.The peri-implant bone losses in the mesial side were (0.67 ±0.35) mm and (0.69 ±0.49) mm, respectively, while in the distal side were (0.73 ±0.31) mm and (0.75 ±0.48) mm, respectively.All these indicators showed no significant difference between the 2 groups ( P>0 .05 ) .Conclusion:Both the cases obtained optimizer results after loading 2 years, and the soft and hard tissues around the implant were very stable , which means that both the protocols can achieve reliable therapeutic effects .If we can handle the indications , immediate implant for anterior teeth shows similar efficacy with delayed implant in the short term .But immediate implant in terms of shortening the course of treatment is clearly superior to delayed implant .

ObjectiveTo study the proper timing for radical hepatectomy after tumour-down-staging with transcatheter arterial chemoembolization for unresectable primary liver cancer.Method This is a retrospective study of 18 patients with unresectable primary liver cancer who received radical liver resection after tumour-downstaging with transcatheter arterial chemoembolization (TACE) from January 2005 to August 2010 at Zhejiang Province People's Hospital Hepatobiliary Surgery Department.The patients received TACE 1 to 3 times (once n=4,twice n=12,and thrice n=2).After tumour-downstaging,radical liver resection was carried out (right hepatectomy,n =10 ; resection of tumour in right liver + resection of right liver metastases,n=2; resection of tumnour in right liver +radiofrequency ablation of right liver metastasis,n=1; right hepatectomy + removal of portal vein tumour thrombus,n=1 ; left hepatectomy + radiofrequency ablation of right liver metastases,n=2 ;Mesohepatectomy,n=1; and left hepatectomy + excision of liver metastasis,n=1).ResultsAfter TACE,the diameter of the primary tumour reduced by over 30％ in 6 patients (6/18,33.3％);10％～30％ in 8 patients (8/18,44.4％),and 10％ in 4 patients (4/18,22.2％).Before TACE,the tumours were not encapsulated in 6 patients (33.3％).After TACE,only 1 patient (5.6％) had the tumour remained unencapsulated.After TACE in 6 patients,the primary tumour shrunk to be within a hemiliver,and ultrasound and CT showed the tumours to have defined borders and they were away from the porta hepatis and major blood vessels.In another 6 patients,there were metastases to the contralateral hemilivers but these tumours had all shrunk in size.Selective vascular inflow and outflow occlusion technique was routinely used for liver resection.ConclusionFor primary liver cancers which are not resectable,TACE should be used first.When the tumours shrink in size,radical resectional surgery should be performed as soon as possible.The surgical technique should follow the following principles:-preserve as much normal liver parenchyma as possible,use selective vascular inflow and outflow occlusion technique to avoid ischaemia/reperfusion injury to the remnant liver,and to reduce haemorrhage.The surgery should be carried out by experienced surgeon.