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Objective:To explore the efficient construction of HSF1 gene knockout mouse model using CRISPR/Cas9 gene editing technology, and to establish the early basis for the mouse model of primary osteosarcoma.Methods:According to exon 9 of HSF1 gene structure, the corresponding GRNA (guideRNA) was selected and screened. Then the transcription template of sgRNA (small guide RNA) was amplified by PCR, and four up stream primers were obtained. Subsequently, sgRNA was transcribed in vitro and screened by Tube Screen platform to screen the sgRNA with effective cutting, and the sgRNA with the highest cutting efficiency was selected from the screening results for subsequent experiments. The transcription template of SPCas9mRNA was amplified by PCR, and then Cas9mRNA was transcribed in vitro. The sgRNA transcribed in vitro and Cas9mRNA were injected into the fertilized eggs of healthy C57BL/6 mice, and the tissue was extracted from the tail of the born mice and identified by PCR sequencing. Heterozygous female mice of F0 generation were selected to mate with wild-type male mice too btain F1 generation off spring. The mutation of gene bases of F1 generation mice was detected by AGAR gel electrophoresis and gene sequencing. The heterozygous male mice of the F1 generation and female mice of the F0 generation were back crossed to obtain the F2 generation daughter mice. The tail tissues were cut and sequenced to obtain the F2 generation homozygous knockout mice. PCR was used to observe the cutting efficiency of sgRNA and the sequencing of rat tail tissue, and SNAPGene software was used for gene sequence alignment to determine the deletion of base fragments.Results:The up stream primers sgRNA-1 Primer-f, sgRNA-2 Primer-f, sgRNA-3 Primer-f, sgRNA-4 Primer-f and down stream primers sgRNA-4 Primer -r were obtained by PCR amplification. After in vitro tran scription and screening of sgrRNA, sgrRNA-1, sgrRNA-2 and sgrRNA-4 had high cleavage efficiency and were selected for subsequent experiments. T7 promoter was added to the 5 'end of Cas9 mRNA, and Cas9 mRNA was obtained by PCR and in vitro transcription kit. Mixed Cas9-sgRNA solution was injected into the fertilized eggs of mice and cultured. The cultured two-cell fertilized eggs were injected into the ampulla of the pseudo pregnant female mice, and the F0 generation mice were obtained successfully. A total of 8 heterozygous mice of F0 generation were obtained by Agar gel electrophoresis. Three heterozygous knockout mice of F1 generation were obtained by breeding the female heterozygous mice of F0 generation with healthy wild-type male mice and PCR and sequencing. Three heterozygous male mice of F1 generation were back crossed with female mice of F0 generation 3 to obtain F2 generation mice. Through the observation of electrophoresis and sequencing results of F2 generation mice, it was confirmed that 7 mice were missing HSF1 base sequence, and the electrophoresis results showed mutant bands and no wild-type bands, which were identified as homozygous. The F2 generation homozygous mice were able to breed stably. As eries of results proved that the HSF1 gene knockout mouse model was successfully established in this experiment.Conclusion:CRISPR/Cas9 technology was successfully used to construct HSF1 gene knockout mouse model, with strong stability and high reproducibility, which laida foundation for further study of HSF1 gene expression products and establishment of mouse model of primary osteosarcoma.
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Objective:To investigate the effect of microRNA-146a (miR-146a) on apoptosis of human retinal microvascular endothelial cells (HRMECs) induced by high glucose and its possible molecular mechanism.Methods:HRMECs were cultured in vitro with 5.5 mmol/L D-glucose in the normal control group and 25 mmol/L D-glucose in the high glucose group for 48 hours, respectively.Normally cultured HRMECs were transfected by miR-146a mimics in the high glucose+ miR-146a mimics group or corresponding mimics control in the high glucose+ mimics control group by lipofection and cultured with 25 mmol/L D-glucose for 48 hours, respectively.Real-time fluorescent quantitative polymerase chain reaction (PCR) was performed to detect the expression level of miR-146a.MTT assay and flow cytometry were used to detect the activity and apoptosis of HRMECs, and Western blot was employed to detect the expression levels of apoptosis-associated protein B-cell lymphoma factor-2 (Bcl-2), Bcl-2 related X protein (Bax) and nuclear factor-κB (NF-κB) signaling-related proteins NF-κB p65 and p-NF-κB p65. Results:The relative expression levels of miR-146a were 1.00±0.10, 0.22±0.02, 0.21±0.02 and 0.88±0.09, and the cell viability was (100.00±10.06)%, (68.41±6.67)%, (67.91±6.74)% and (90.46±8.97)%, and the apoptosis rates were (3.11±1.02)%, (27.28±3.56)%, (27.44±4.03)% and (7.29±2.11)% in the normal control group, high glucose group, high glucose+ mimics control group and high glucose+ miR-146a mimics group, respectively.The relative expression levels of miR-146a and the cell viability were significantly lower, and the cell apoptosis rate was significantly higher in the high glucose group than those in the normal control group, with statistical significant differences (all at P<0.05). The relative expression levels of miR-146a and the cell viability were significantly higher, and the cell apoptosis rate was significantly lower in the high glucose+ miR-146a mimics group than those in the high glucose group and the high glucose+ mimics control group, and the differences were statistically significant (all at P<0.05). The relative expression levels of Bax and p-NF-κB p65 protein were significantly higher, the relative expression level of Bcl-2 protein was significantly lower in the high glucose group than those in the normal control group, showing statistically significant differences (all at P<0.05). The relative expression levels of Bax and p-NF-κB p65 protein were significantly lower, and the relative expression level of Bcl-2 protein was significantly higher in the high glucose+ miR-146a mimics group than those in the high glucose group and the high glucose+ mimics control group, and the differences were statistically significant (all at P>0.05). There was no significant difference in the relative expression of NF-κB p65 protein among the groups ( F=0.106, P=0.955). Conclusions:Overexpression of miR-146a may inhibit the apoptosis of HRMECs induced by high glucose, and its mechanism may be related to the inhibition of NF-κB signaling pathway activation.
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Objective:To investigate the effect of dexmedetomidine combined with flurbiprofen axetil on pain and cognitive function of patients after thyroidectomy.Methods:A total of 80 patients with thyroid gland surgery in the People's Hosptial of Shouguang from January 2016 to December 2017 were selected and randomly divided into four groups according to the digital table: saline control group(C), dexmedetomidine group(D), flurbiprofen axetil group(F), dexmedetomidine combined with flurbiprofen axetil group(DF), with 20 casess in each group.The extubation time and wake-up time of the four groups were observed.The VAS scores were used to assess the analgesic effect after surgery, and the Riker sedation scale was used to assess postoperative anxiety.Results:The extubation time of the C, D, F and FD group were (16.66±3.37)min, (24.63±2.80)min, (14.55±3.74)min, (26.22±3.45)min, respectively, which of group D and group DF was significantly longer than that in group C and group F( F= 4.59, P<0.01). The recovery time of group D[(17.98±2.54)min] and group DF[(17.84±3.63)min] was significantly longer than that of group F[(10.92±2.21) min] and group C[(9.95±2.16)min] ( F=5.01, P<0.01). The VAS scores of group DF at 30 min, 3 h, 6 h and 24 h after recovery were significantly lower than those of group F and group D ( F=4.53, 4.59, 4.61, 4.35, all P<0.01). The Rick sedation scores of C, D, F and FD groups were (5.28±1.81)points, (4.20±1.56)points, (4.24±2.01)points and (3.76±1.43)points, respetively, which of group DF was significantly lower than those of the other three groups( F=3.01, P<0.01). The MMSE scores of cognitive function in group DF at 3 h, 1 d and 3 d after surgery were significantly higher than those in group C ( F=5.89, 4.95, 4.56, all P<0.01). Conclusion:The combined use of dexmedetomidine and flurbiprofen axetil significantly reduces pain and relieves postoperative cognitive impairment after thyroid surgery.
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Objective@#To investigate the effect of dexmedetomidine combined with flurbiprofen axetil on pain and cognitive function of patients after thyroidectomy.@*Methods@#A total of 80 patients with thyroid gland surgery in the People's Hosptial of Shouguang from January 2016 to December 2017 were selected and randomly divided into four groups according to the digital table: saline control group(C), dexmedetomidine group(D), flurbiprofen axetil group(F), dexmedetomidine combined with flurbiprofen axetil group(DF), with 20 casess in each group.The extubation time and wake-up time of the four groups were observed.The VAS scores were used to assess the analgesic effect after surgery, and the Riker sedation scale was used to assess postoperative anxiety.@*Results@#The extubation time of the C, D, F and FD group were (16.66±3.37)min, (24.63±2.80)min, (14.55±3.74)min, (26.22±3.45)min, respectively, which of group D and group DF was significantly longer than that in group C and group F(F= 4.59, P<0.01). The recovery time of group D[(17.98±2.54)min] and group DF[(17.84±3.63)min] was significantly longer than that of group F[(10.92±2.21) min] and group C[(9.95±2.16)min] (F=5.01, P<0.01). The VAS scores of group DF at 30 min, 3 h, 6 h and 24 h after recovery were significantly lower than those of group F and group D (F=4.53, 4.59, 4.61, 4.35, all P<0.01). The Rick sedation scores of C, D, F and FD groups were (5.28±1.81)points, (4.20±1.56)points, (4.24±2.01)points and (3.76±1.43)points, respetively, which of group DF was significantly lower than those of the other three groups(F=3.01, P<0.01). The MMSE scores of cognitive function in group DF at 3 h, 1 d and 3 d after surgery were significantly higher than those in group C (F=5.89, 4.95, 4.56, all P<0.01).@*Conclusion@#The combined use of dexmedetomidine and flurbiprofen axetil significantly reduces pain and relieves postoperative cognitive impairment after thyroid surgery.
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Objective:To propose and verify a surgical classification system for the axial primary malignant and aggressive benign tumor.Methods:The CZH surgical classification system was originally developed for the axial primary malignant and aggressive benign tumor. The CZH surgical classification system includes seven types, according to the anatomic features and the extension of tumor violation. A total of 136 patients (79 males and 57 females) with axial primary malignant and aggressive benign tumor from multiple tertiary centers who received surgery from July 2006 to July 2019 were included. The average age was 44.40±17.55 years (8-83 years) old. There were 99 malignant tumors and 37 aggressive benign tumors included. The number of patients with each classification was presented as followed, Type I 13, Type II 15, Type IIIa 3, Type IIIb 20, Type IVa 43, Type IVb 12, Type Va 21, Type Vb 3, Type VI 2, Type VIIa 3 and Type VIIb 1. Surgical procedures were selected according to different types in classification. The inter- and intra-observer consistencies were evaluated by the Kendall's W test. The VAS, Frankel score, overall survival and recurrence free survival were recorded during the follow-up. Results:The inter- and intra-observer consistent coefficient was 0.973 and 0.996, respectively ( P<0.05). The single posterior approach was adopted for the Type II tumors. Other patients underwent surgery by the combined antero-posterior approach. The majority in anterior approach (113 cases) was the modified submandibular approach. The reconstruction modes included anterior "T" shape titanium mesh (112 cases) or the 3D printed prothesis (7 cases) combined with the posterior occipto-cervical fusion (92 cases) or the pedicle screw system (44 cases). The average surgical duration and the volume of intraoperative bleeding was 348.40±136.14 min (60-760 min) and 1 225.69±859.40 ml (80-4 000 ml), respectively. The operation duration and volume of intraoperative bleeding among each type were with statistical difference. The patients with Type IV, V tumors had longer operation duration than those with Type II tumors. Those with Type V and VII tumors had longer operation duration than those with Type I tumors. The patients with Type V tumors had more intraoperative bleeding than those with Type I-IV tumors. The average preoperative VAS score was 4.15±2.25 and then was reduced significantly to 0.62±0.71 and 0.38±0.59 at one and three months after operation, respectively. The Frankel score was also significantly ameliorated at one and three months postoperatively. There were 22 postoperative complications (16.2%). The complications included cerebral spinal fluid leak (12.5%), dysphagia and/or dysphonia (7.4%), dyspnea (5.1%), wound infection (3.7%), wound hemorrhage (2.2%) and pharyngeal dehiscence (1.5%). The incidence of postoperative complication was 25.9% in Type IV-VII tumors, while 11.8% in Type I-III tumors. Conclusion:CZH surgical classification system was verified with high observer consistency. This classification system could assist surgeons to select proper surgical approaches, resection modes and reconstruction modes, and thus ensure the safety of surgery and reduce the recurrence. The tumors in Type IV, V and VII may be with more challenging for surgeons. The incidence of postoperative complication in Type IV-VII tumors may be higher than that in Type I-III tumors.
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Objective To study the feasibility and safety of total en bloc spondylectomy (TES) for bone tumors of the fourth lumbar spine and evaluate the clinical outcomes.Methods From March 2011 to December 2013,21 patients undergone total en bloc spondylectomy in posterior-only approach were retrospectively reviewed.The patients included 9 males and 12 females,with a mean age of 47.1± 15.6 years old (range,15-71 years old).This series included 12 cases of primary bone tumors and 9 cases of solitary metastases.Preoperative evaluation according to clinical,imaging and pathologic features was performed meticulously to select patients.The length of surgery,estimated blood loss,surgical margins,instrumentation failure,perioperative complications,Frankel scale,visual analogue scale (VAS) for pain,local control rate and overall survival were reviewed and analyzed.Results Total en bloc spondylectomy was performed successfully in all patients.Average operative time and estimated blood loss were 297.6±44.6 min (range,225-420 min) and 2 247.1±904.5 ml (range,900-4100 ml),respectively.The mean follow-up time was 50.4± 17.1 mons (range,24-79 mons).All patients encountered nerve roots stretch and 5 patients (23.8％) showed lower extremeties neurological dysfunction.All of them improved in 2-4 weeks postoperatively and recovered completely at 6-month follow -up.Cerebrospinal fluid leak was found in 4 patients (19.0％).The VAS score was 1.5±1.4 at post-operation,which was significantly lower than the 6.2± 1.6 in average at operation (P=0.008).Three patients with metastatic tumors died during the follow-up.Titanium mesh cage subsidence was observed in 7 patients (33.3％).No implant failure was occurred during the follow-up.Conclusion Total en bloc spondylectomy for tumors of the fourth lumbar spine in a posterior-only approach is feasible.However,there are many intraoperative neurological complications and the indications for TES are extremely limited.
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Background Studies determined that blue light exposure causes apoptosis of human retinal pigment epithelial (RPE) cells,but its mechanism is still below understood.Objective The aim of this study was to investigate whether or how mitochondrial apoptotic pathway is involved in blue-light induced apoptosis of human RPE cells in vitro.Methods Human RPE cells were isolated from fresh donor eyes and primarily cultured and passaged.The cells were identified with keratin antibody by immunochemistry.Then the cells were the non-light exposed group,simple light-exposed group,light-exposed+nifedipine group,light-exposed+calphostin C group and the light-exposed+phorbol myristate acetate (PMA) group.Human RPE cells in light-exposed group were consequently cultured for 24 hours following the exposure of (2 000±500)lx blue-light for 6 hours,and then the expression levels of bax,bcl-2,bcl-xl in the cells were detected by Western blot to evaluate the effect of blue light on the apoptosis.The cells in the light-exposed+nifedipine group,light-exposed+calphostin C group and the light-exposed+PMA group were treated with the corresponding drugs 1 hour prior to light irradiation and sequently received 6-hour light irradiation and 48-hour culture.The expression of caspase-9 protein in the cells were assayed with Western blot to assess the influence of Ca2+ channel and protein kinase C (PKC) pathway on mitochondria of RPE cells.Results Cultured cells grew well with visible pigment in cytoplasm.The cells showed the positive response for keratin and presented a cobblestone-like appearance.The expression bands of bax,bcl-2 and bcl-xl proteins were clearly visible at the molecular weight of 23 000,26 000 and 30 000 in both non-light exposed group and the simple light-exposed group,and the absorbance values of the cells to bax were elevated,while the absorbance values to bcl-2 and bcl-xl were declined in the simple light-exposed group compared with the non-light exposed group (t =-4.409,P =0.012 ;t =7.575,P =0.002 ; t =6.068,P =0.004).Compared with the non-light exposed group,the absorbance values of caspase-9 were significantly raised in the simple light-exposed group,light-exposed+calphostin C group and the lightexposed+PMA group (P=0.005,0.002,0.000),but no significant difference between the non-light exposed group and light-exposed+nifedipine group (P=0.191).Compared with the simple light-exposed group,the expression level was considerably higher in the light-exposed + PMA group (P =0.005) ; while that in the light-exposed + nifedipine group or light-exposed+calphostin C group was not significantly different (P=0.057,0.643).Conclusions Blue light exposure induces apoptosis of RPE cells by up-regulating the expressions of bax and caspase-9 proteins and down-regulating the expressions of bcl-2 and bcl-xl.The mitochondrial apoptosis pathway and PKC pathway participate in blue-light induced apoptosis of human RPE cells in vitro.
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Objective To investigate the application value of continuous irrigation and vacuum suction by subcutaneous drainage tube for prevention of abdominal type Ⅳ incision infection.Methods A prospective,single-blind, randomized, controlled study was conducted based on the clinical data of 123 patients with abdominal type Ⅳ incision infection who were admitted to the First People's Hospital of Foshan between January 2008 and July 2014.Patients were divided into the experimental group and the control group based on the random number table and received open surgery.Patients in the experimental group were placed subcutaneous drainage tube with postoperative continuous irrigation and vacuum suction, while patients in the control group adopted the method of traditional abdominal closure without subcutaneous drainage tube.The levels of preoperative hemoglobin (Hb) and albumin (Alb), severity grading according to the American Society of Anesthetheologists (ASA), levels of Hb and Alb at postoperative day 1, 3, 7 were recorded and postoperative incision infection and bacteria culture were observed.Patients received bi-weekly regular return visit by outpatient evamination after discharged up to 3 months after suture removal.Measurement data with normal distribution were presented as x-± s and comparison between groups was analyzed by t test.Measurement data with skew distribution were presented as M (Qn) and comparison between groups were analyzed by rank sum test.Repeated measures data were analyzed by the repeated measures ANOVA.Count data were analyzed by the chi-square test.Results One hundred and twenty-three patients were screened for eligibility, and 65 were allocated into the experimental group and 58 into the control group.The levels of Hb and Alb at postoperative day 1, 3, 7 were (111 ± 15) g/L, (107 ± 18) g/L, (108 ± 13) g/L and 30 g/L (26 g/L,32 g/L), 31 g/L(28 g/L,33 g/L), 35 g/L(32 g/L,37 g/L) in the experimental group and (112 ± 13)g/L, (106 ±16)g/L, (106 ± 12)g/L and 30 g/L(25 g/L,32 g/L), 32 g/L(29 g/L,33 g/L), 37 g/L (32 g/L,38 g/L) in the control group, with no significant difference in the changing trends of the above indexes between the 2 groups (F =0.124, 0.007, P ＞ 0.05).There were 4 patients with incision infection in the experimental group and 12 patients in the control group, showing a significant difference (x2=2.723, P ＜ 0.05).The infections occurred at postoperative 4-6 days confined to subcutaneous tissues and unreached to muscular and below layers, and then were cured by incision open drainage without recurrence.Patients without incision infection were removed the stitches at postoperative 7-9 days.Incision bacteria cultures showed that Escherichia coli was detected in 8 cases (including 1 complicated with Enterococcus faecalis and 1 complicated with Klebsiella pneumonia and Pseudomonas aeruginosa), Pseudomonas aeruginosa in 1 case, Klebsiella pneumonia in 2 cases, Acinetobacter baumanii in 1 case, Enterobacter cloacae in 1 case and no bacteria in 3 cases.All the patients were followed up for 3 months after incision healing and survived well without recurrence and complications such as incision split and incision fistula.Conclusion Continuous irrigation and vacuum suction by subcutaneous drainage tube can be operated easily and effectively reduce the postoperative infection rate in abdominal type Ⅳ incision.
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Objective To evaluate the mechanical stability of alternative reconstruction methods after total en bloc spon?dylectomy in the lower lumbar spine. Methods Eight adult fresh cadaveric lumbosacral spines (L1-S1) were adopted. Total en bloc spondylecotmy of the L4 vertebra was performed after intact testing. Four designed reconstruction samples were tested for the range of motion (ROM) of the spine:1) expandable artificial vertebral body and short posterior instrumentation at L3-L5 (SP), 2) ex?pandable artificial vertebral body and short posterior instrumentation with additional anterolateral fixation at L 3-L5 (ASP), 3) ex?pandable artificial vertebral body and multilevel posterior instrumentation at L2-S1 (MP), 4) expandable artificial vertebral body and multilevel posterior instrumentation with additional anterolateral fixation at L2-S1 (AMP). Nondestructive biomechanical test?ing was performed on each construct under loading control. The ROM for each construct was obtained by applying pure moments in flexion, extension, lateral bending, and axial rotation. Results In flexion, extension and lateral bending, the ROM of all the re?constructed constructs significantly decreased compared to the intact. The ROM of specimens with anterolateral fixation was less when compared to the ones without additional fixation. In lateral bending, MP (L:1.87° ± 0.32° , R:1.97° ± 0.33° ), ASP (L:1.89° ± 0.37°, R:2.08°±0.36°) and AMP (L:1.32°±0.29°, R:1.61°±0.33°) provided significantly less ROM than the SP (L:3.14°±0.35°, R:3.44°±0.34°). In axial rotation, the ROM of ASP (L:4.21°±0.58°, R:4.02°±0.59°) and AMP (L:3.56°±0.55°, R:3.52°±0.48°) was significant decreased when compared to the intact state (L: 7.47° ± 1.00° , R:7.57° ± 0.84° ). MP (L:6.33° ± 0.71° , R:5.88° ± 0.62°), ASP and AMP showed significantly less ROM than the short posterior fixation (L:9.28°±1.01°, R:9.48°±0.98°). AMP sig?nificantly decreased the ROM compared to MP. Conclusion After total en bloc spondylectomy of lower lumbar, long segmental fixation provided more stability to the construct than the short one. Compared to posterior fixation, circumferential fixations showed a higher stability. In contrast, multilevel segmental instrumentation with circumferential fixation did not provided more stability than the short constructs.
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Objective To investigate the safety and feasibility of modified hand-assisted laparoscopic surgery (MHALS) in the abdominal surgery.Methods The clinical data of 8 patients who underwent long-sleeved MHALS at the First People's Hospital of Foshan between September 2014 and January 2015 were retrospectively analyzed.Among the 8 patients,right liver cancer with intrahepatic metastasis was found in 2 patients,left liver cancer in 1 patient,hepatic peripheral nerve sheath tumor in 1 patient,left retroperitoneal leiomyosarcoma in 1 patient,extra-and intra-hepatic cholangiolithiasis in 1 patient,choledochocyst in 1 patient and ampulla cancer in 1 patient.Laparoscopic protection sleeve went through the middle of incision-retractor,and then wrapped around it about 10 cm.Incision-retractor was fixed at the abdominal incision firstly,laparoscopic procedures were performed when the wrist of assisted hand was bound and fixed by the distal of sleeve.The patients were followed up by outpatient examination and telephone interview till March 2015.Results All the 8 patients underwent successful MHALS,including 1 of right hemihepatectomy in situ,1 of ligation of right portal vein + left liver split (the patient gave up two-stage operation due to intractable ascites and elevated bilirubin),1 of hepatic left lateral lobectomy (the patient underwent hemostatic sutures in open surgery due to hemorrhage of liver's cutting surface),1 of hepatic peripheral nerve sheath tumor resection,1 of left retroperitoneal leiomyosarcoma resection,1 of choledocholithotomy + left hepatectomy + cholecystectomy + T tube drainage,1 of choledochocyst + biliary enteric drainage and 1 of pancreaticoduodenectomy.Four patients had assisted incision of 4 cm,and another 4 patients of 7 cm.Eight patients were followed up for a median time of 3 months (range,2-7 months).The patient who received ligation of right portal vein + left liver split died at postoperative month 3,and the others didn't have recurrence of tumor or lithiasis.Conclusion The MHALS is safe and feasible in the abdominal surgery.
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Objective To explore the application value of hemi-hepatic blood flow occlusion through descending hilar plate in laparoscopic anatomic hepatectomy.Methods The clinical data of 15 patients who underwent laparoscopic anatomic hepatectomy by hemi-hepatic blood flow occlusion using descending hilar plate technique at the First People's hospital of Foshan between August 2012 and May 2014 were retrospectively analyzed.The hilar plate was bluntly dissected to expose the left and right Glissonean pedicles.Either side of Glissonean pedicle was tied up with a turnable aspirator with a cotton rope or shoelace and then bypassed the back of hilar plate.Anatomic hepatectomy was performed when hemi-hepatic blood flow was occluded.The follow-up by telephone interview and outpatient examination was done till October 2014.Results Among the 15 patients,the conversion to open surgery was done in 1 patient,Pringle maneuver in 1 patient,and hemi-hepatic blood flow occlusion by descending the hilar plate in 14 patients.Thirteen patients received succesfully laparoscopic anatomic hepatectomy by hemi-hepatic blood flow occlusion using descending hilar plate technique,including 4 of left hemihepatectomy,4 of left lateral lobectomy,2 of right hemihepatectomy,1 of right posterior lobectomy,1 of segment Ⅳ hepatectomy and 1 of segment Ⅵ hepatectomy.Bile duct exploration was applied to 4 patients with left hepatic duct stones and T-tube was placed in 2 patients.Nine and 4 patients received left and right hemi-hepatic blood flow occlusion,respectively.The operation time,mean volume of intraoperative blood loss and time of hemi-hepatic blood flow occlusion in 13 patients were (196 ±63)minutes,320 mL (range,50-1 200 mL) and (51 ± 20)minutes,respectively.The time of descending the hilar plate in 14 patients was (10 ±4)minutes.Among the 13 patients,bile leakage was detected in 1 patient with a maximum volume of drainage of 120 mL/day,liver wound bleeding in 1 patient with a volume of abdominal bloodstained drainage of 400 mL at postoperative day 2.Two patients were cured by conservative treatment,and no liver failure and perioperative death were occurred.The duration of hospital stay was (6.9 ± 2.4)days.Among the 15 patients,2 patients were loss to follow-up and other patients were followed up for 5-26 months with good survival,1 patient died.Conclusion Hemi-hepatic blood flow occlusion through descending hilar plate in laparoscopic anatomic hepatectomy is safe and feasible.
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group was significantly longer than that in the low MELD score group (P <0.05).And there was no significant difference in the length of hospital stay,mortality during hospitalization,incidence of early complications, follow-up time and overall survival rate between two groups (all in P >0.05).The peak level of aspartate aminotransferase (AST)in the high MELD score group was significantly higher compared with that in the low MELD score group (P <0.05 ).The incidence of bile leakage,abdominal abscess and liver dysfunction significantly differed between two groups (all in P <0.05 ).Conclusions It is a safe and short-term efficacious approach to perform liver transplantation with organs obtained from Chinese donation after citizens’ death in patients with high MELD score liver recipients.
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Objective To discuss the classification and treatment of the degloving hand.Methods All 68 degloving hand treated between January, 2005 and December, 2014 in our department were reviewed.The cases were divided into three types according to the extent of the injury.Type Ⅰ the degloving tissue had enough blood supply which was debrided and stitched.Type Ⅱ the degloving tissue had less or no blood supply which was revascularized with replantation or vein anastomosis, some degloving palm or dorsum resurfaced the wound by the degloving skin graft.Type Ⅲ the degloving tissue had no blood supply or severely damaged,the tissue defect should be resurfaced with other tissue reconstruction.Type Ⅱ and Ⅲ were further divided into subtype A, B and C depending on the degloving scope and location, each subtype has its own surgical procedure.Results The total survival rate was 83.8％.The survival rate decreased with the increasing extent of the injury and the increasing degloving scope.Conclusion Classification of the hand degloving injuries depending on the extent of the injury, the degloving scope and location is useful for the assessment of injury and the selection of proper operation procedures.
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Objective To investigate the effects of ischemic preconditioning on the cholesterol content and the activity of Na+-K+-ATPase of hepatocytes following cold preservation in rats.Methods Twenty-five rats were randomly divided into five groups,including control group (C),cold preservation group (Ⅰ),ischemic preconditioning group (ⅠP),atorvastatin (30 μmol/L) treatment group (A30),and atorvastatin (100 μmol/L) treatment group (A100).The cholesterol content and the activity of Na+ -K+ -ATPase were assessed.Results The cholesterol contents on the rat liver tissue cell membrane in the C group,Ⅰ group,ⅠP group,A30 group and A100 group were (310.4 ± 27.5),(187.7±13.1),(394.3±25.9),(201.8±14.6) and (122.6±7.7) nmol/mg protein,and activity of the Na+ -K+ -ATP enzyme was (46.55 ± 3.20),(27.4 ± 2.81),(52.71 ± 3.02),(30.67 ±2.78) and (19.64 ± 2.11) μmol Pi/hr mg protein,respectively (P＜0.05).There was no significant difference in the plasma membrane phospholipid content among the five groups (P＞0.05).Conclusion Reduction of cholesterol content and Na+ K+ -ATPase activity on the liver cytoplasmic membrane is one of the factors causing donor liver cold preservation injury,but ischemic preconditioning can significantly improve cell membrane Na+ -K+ -ATPase activity and increase cytoplasmic membrane cholesterol content. Use of atorvastatin statins can reduce cytoplasmic membrane cholesterol synthesis,and significantly decrease Na+ -K+ -ATPase activity,thereby alleviating the donor liver cold preservation injury.
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Objective To investigate procedure and therapeutic effect of resection and reconstruction for axis tumors through the sub mandible approach. Methods Between December 2004 to June 2010,17 patients with axis neoplasm underwent tumor resection and antero-posterior reconstruction through the combined the sub mandible-inner sternocleidomastoid muscle (SMIS) approach and posterior approach. Tumor lesions involved C2 in 11 cases, C2-3 in 4, C2-4 in 2. Eight cases involved vertebral body, and 9 involved both vertebral body and element. Fourteen primary lesions including 4 giant cell tumors, 4 plasmocytomas, 2 chordomas, 2 eosinophilic granulomas, 1 hemangiopericytomas and 1 lymphoma, and 3 metastatic lesions were involved in this study. Three types of reconstruction in upper cervical spine including titanium mesh plus vertically placed titanium plate, titanium mesh plus obliquely placed titanium plate and trimmed titanium mesh alone, were adopted after anterior tumor resection, and then posterior tumor resection and reconstruction were performed. Results All patients experienced pain relief and neurological improvement after surgery. Except for one incidence of screw pull-out which was corrected by a revision surgery, solid fusion was achieved in all patients. A follow-up period of 6 months to 6 years was available for this study. One patient died of cerebral infarction 9 months postoperative. Two patients with chordoma relapsed 13 months and18 months postoperative, respectively, of whom one died of high plegia and respiratory failure, and the other was alive with disease. Two patients with metastasis died of multiple remote metastases 12 months and 18 months postoperative, respectively. Conclusion Through the SMIS apporach, a satisfactory exposure can be obtained for axis tumor resection and reconstruction. Anterior reconstruction of upper cervical spine after tumor resection can be achieved with internal fixation system of cervical spine, which can improve intraopera-tive safety. The combined anterior reconstruction and posterior occipito-cervcial fixation can provide immediate stability, and benefit maintaining stability of upper cervical spine.
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This paper is to establish a three-dimensional finite element model (3D-FEM) of pelvic floor levator ani muscles in an old healthy women. We acquired the image data of the pelvic bones and pelvic floor muscles from CT and MRI scanning in a non-pregnant old healthy female volunteers. The 3-D reconstruction and mesh optimization of the whole pelvic bones and muscles with application of image processing software Mimics12.0 and Geomagic9.0 were obtained. Then we built the 3D-FEM of the musculoskeletal system of the pelvic bones and levator ani muscles with Ansys11.0 software. We obtained an accurate 3D-FEM of pelvic bones and levator ani muscles in the older healthy woman. The results showed that it was reliable to build 3D-FEM with CT and MRI scanning data and this model could vividly reflect the huge space anatomy of the real pelvic floor levator ani muscles. It avoids the defects to gain the model from the body of anatomical specimens in the past. The image data of model are closer to vivisection, and the model is more conducive to the latter finite element analysis.
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Female , Humans , Middle Aged , Finite Element Analysis , Imaging, Three-Dimensional , Magnetic Resonance Imaging , Muscle, Skeletal , Diagnostic Imaging , Pelvic Floor , Diagnostic Imaging , Tomography, X-Ray Computed , MethodsABSTRACT
Objective To evaluate the efficacy of liver grafts with warm ischemia and with different cold preservation time in liver transplantation.Methods The clinical data of 154 patients who received liver transplantation at the First Affiliated Hospital of Sun Yat-sen University from January 2006 to December 2007 were retrospectively analyzed.The warm ischemia time of the liver grafts obtained from the non-heart-beating donors was within 10 minutes.According to cold perservation time of the liver grafts,patients were divided into 3 groups:the cold preservation time of the liver grafts was within 8 hours,8-12 hours and above 12 hours in group I(n=58),group Ⅱ(n=62)and group Ⅲ(n=34),respectively.The peak level of alanine aminotransferase(ALT),primary graft dysfunction(PGD)after liver transplantation,acute rejection response,biliary complications,vessel complications,perioperative infections and the survival of liver grafts and recipients among the 3 groups were analyzed via chi-square test,t test and variance analysis.Results No PGD was detected in the 3 groups after liver transplantation.All patients were followed up for 8-32 months.The peak level of ALT,incidence of infection and biliary complication,survival of liver grafts and recipients were(482±357)U/L,12%(7/58),12%(7/58),86%(50/58)and 88%(51/58)in group Ⅰ,and were(1274±608)U/L,29%(10/34),26%(9/34),68%(23/34)and 71%(24/34)in group Ⅲ,with significant difference between the 2 groups(t=5.23,X~2=4.28,6.77,4.51,4.28,P<0.05).The peak level of ALT in group Ⅱ was(953±424)U/L,which was significant higher than(482±357)U/L in group Ⅰ(t=4.76,P＜0.05).Conclusions Liver grafts with a warm ischemia time shorter than 10 minutes could tolerate the injury caused by cold preservation with the maximum time of 12 hours.The incidences of biliary complications and postoperative infections are significantly increased and the survivals of liver grafts and recipients are decreased when the cold preservation time exceeds 12 hours.
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BACKGROUND:Nowadays,liver grafts for transplantation are clinically sourced from non-heart-beating donors.Moreover,there is still no uniform determination of safe time limit points for non-heart-beating donor liver in warm ischemia and cold preservation.OBJECTIVE:To evaluate the application safety and curative effects of warm ischemia liver graft affected by varied cold ischemia time (CIT) in liver transplantation.DESIGN,TIME AND SETTING:A randomized controlled observation was performed in the Organ Transplantation Center,First Affiliated Hospital of Sun Yat-sen University between January 2006 and December 2007.PARTICIPANTS:154 cases who underwent non-heart-beating liver transplantation were included in this study.All liver graftsinvolved had a warm ischemia time(WIT) less than 10 minutes.METHODS:All cases were assigned into 3 groups according to CIT:group Ⅰ (CIT＜8 hours,n=58),group Ⅱ (CIT 8-12hours,n=62),and group Ⅲ (CIT 13-16 hours,n=34).The liver grafts were randomly allocated for the patients.Following liver transplantation,the same immunosuppression protocol was employed for each group.MAIN OUTCOME MEASURES:Following surgery,peak level of alanine aminotransferase (ALT),primary graft dysfunction (PGD) after liver transplantation,acute rejection response,biliary complications,vessel complications,perioperative infections,graft and recipient survival rate were compared among 3 groups.RESULTS:Follow-up time was 8-32 months.No PGD was detected in all 154 cases.Group Ⅱ showed postoperative ALT peak levels significantly higher than group Ⅰ (P＜ 0.05).There was no significant difference between groups Ⅰ and Ⅱin terms of acute cellular rejection,perioperative infection,biliary complication,vessel complication,graft survival rate,and recipient survival rate (P＞0.05).Compared with group Ⅰ,the group Ⅲ exhibited significantly increased postoperative ALT peak level,biliary complications,and perioperative infections,and significantly decreased graft and recipient survival rate (P＜0.05).CONCLUSION:Non-heat-beating-liver grafts with less than 10 minutes of warm ischemia can tolerance 12 hours of cold ischemia.More than 12 hours,postoperative liver transplantation complications ascend and,contradictorily,graft and recipient survival rate descend.