To produce monoclonal antibody (mAb) specifically against human thrombomodulin (hTM), an immune-tolerizing procedure was employed to generate monoclonal antibodies specific to hTM. Female BALB/c mice were first immunized with CHO cells following at 10 min, 24 h, 48 h by intraperitoneal injection of different doses of cyclophosphamide (CP) 2 times at an interval of 2 weeks, thereby tolerizing the mice to common epitopes shared between CHO and CHO-TM5 cells. Subsequently the selected mice with the lowest titer of serum polyclonal antibody by cellular enzyme-linked immunoabsorbent assay (CELISA) were immunized with CHO-TM5 cells, which have stable high level expression of hTM, to produce antibodies specific to hTM 3 times at an interval of 2 weeks. On the third day after the third immunization, mouse with the highest titer of serum polyclonal antibody was sacrificed and spleen cells were harvested to prepare hybridoma cells with SP2/0 cells at the ratio of 10 to 1. Hybridoma cells were then cultured at 96 well plates for screening with CELISA. To improve probability to obtain specific mAb, CELISA was applied twice. The first CELISA was done with polyethylene ELISA plate with a monolayer of CHO-TM5 cells. The positive clones from the first screen were then selected by reacting with similar screening ELISA plate but having CHO cells monolayer instead. Only clones that were positive for the first screening and negative for the second screening were kept, and called as CHO-TM5 +CHO- hybridoma cells. BALB/c mice were intraperitoneally injected with the selected hybridoma cells. Ascites were collected and monoclonal antibodies were purified using FPLC, and its Ig class, subclass, and titer were then determined respectively. The specificity of the yielded mAb was identified with CELISA, flow cytometry, ABC immunohistochemistry and immunoblotting. Detection of CELISA showed that 100 mg/kg dose of CP could tolerize the mouse to common epitopes shared between CHO and CHO-TM5 cells. And CELISA also discovered that all hybridomas positive for CHO-TM5 cells were negative for CHO cells. Five lines of positive hybridoma cells had been obtained altogether and 2F7 was selected randomly for next investigation. The Ig subclass of the mAb 2F7 was IgG1 and the titer of ascitic mAb was 1?10-6. Furthermore, the content of ascitic mAb was 19.56 g/L and chromosome numbers is 98. Flow cytometry, CELISA and Western blotting assays demonstrated that mAb 2F7 could specifically recognize hTM expressed on CHO-TM5 and human umbilical vascular endothelial cells (HUVEC). Meanwhile, the tissue specificity of mAb 2F7 was also identified by immunohistochemical ABC staining. On the other hand, Western blotting assays indicated that mAb 2F7 could recognize the antigen protein with 105 ku molecular mass under reduction condition. Moreover, the dissociation constant of mAb 2F7, 1.22? 10-9 mol/L, indicated the affinity higher than some others. The results suggest that the immunotolerizing protocol provides a convenient general method for producing antibodies specific to desired protein isoforms. mAb 2F7 can specifically recognize the natural hTM expressed mainly on vascular endothelial cells, which will potentially useful for investigating the functions and clinic values of hTM.

Objective:To investigate the application valve of bFGF to improve the viability of subdermal vascular network flap.Methods:Four white healthy pigs were used,each pig had 6 SVNF on back,16cm?4cm,totally 24 flaps were divided randomly into 2 groups:The bFGF groups and the control groups.On postoperative day 6,we observed the appearance of the flaps.The survival area of each flap was measured and the flap was harvested for histological analysis and the density of angiogenesis was also examined.Results:On postoperative day 6,the survival rates of the bFGF group and the control group were (81.2?1.7)% and (66.2?1.8)% respectively,the angiogenesis density of the middle and the distal segment in the bFGF group was (16.7?6.0)/mm2,(29.7?5.5)/mm2,(14.9?6.0)/mm2 respectively,while (16.6?4.0)/mm2,(21.4?5.2)/mm2,(3.5?5.1)/mm2 respectively in the control group.The bFGF group was superior to the control group in the appearance and histological analysis of the flaps.Conclusion:The results suggest that the injection of bFGF locally can promote the survival rate by accelerating the blood circulation of the SVNF’ bottom and edge,shorten the division time of SVNF and broaden the ratio of length and width.

OBJECTIVETo discuss the operation method and characteristic of correcting upper eyelid depression by transposition of orbital septum fat.

METHODSDuring the double eyelid surgery, we set.the lateral orbital septum fat completely free, while the bottom is still connected with the middle orbital septum fat. We separate a tunnel from the middle to the inner side in orbital septum, and the separated orbital septum fat is transposed to the inner side of orbital septum by the tunnel with suturing fixation.

RESULTSFrom March 2008 to October 2013, 51 cases with upper eyelid depression were treated successfully. Patients were followed up for 3 months to 3 years (average, 7. 5 months) with sustained aesthetic results.

CONCLUSIONSOrbital septum fat transposition can successfully correct the upper eyelid depression. It should become a regular procedure in blepharoplasty.

Adipose Tissue ; transplantation ; Blepharoplasty ; methods ; Esthetics ; Eyelids ; abnormalities ; surgery ; Humans ; Orbit

Objective:To explore intrinsic mechanisms underlying the inhibitory effect of resveratrol on neovascularization in cutaneous squamous cell carcinoma from the perspective of ubiquitin/ubiquitin-like protein modification balance.Methods:The human cutaneous squamous cell carcinoma cell line A431 was used as the research object. Cultured A431 cells at exponential growth phase were divided into 3 groups (control group, 50 μmol/L resveratrol group, and 100 μmol/L resveratrol group) to be cultured with mediums containing 0, 50, and 100 μmol/L resveratrol, respectively. Cell proliferation activity was assessed by the 3- (4,5) -dimethylthiazol (-z-y1) -2,5-di-phenytetrazoliumromide (MTT) assay after 48-hour culture; the vasculogenic mimicry formation assay was performed to evaluate the vasculogenic mimicry formation ability of A431 cells after 12-hour treatment with resveratrol; Western blot analysis was conducted to detect the relative protein expression levels of ubiquitin, small ubiquitin-related modifier-1 (SUMO1), hypoxia-inducible factor 1α (HIF-1α), and vascular endothelial growth factor receptor (VEGFR) in different groups after 48-hour treatment with resveratrol. Then, 24 8-week-old BALB/c male thymectomized mice were randomly and equally divided into 3 groups to be subcutaneously inoculated with A431 cells in the inguinal region, followed by intraperitoneal injections of 1 mg/kg or 2 mg/kg resveratrol (1 mg/kg or 2 mg/kg resveratrol group), or the same volume of physiological sodium chloride solutions (control group) ; the intraperitoneal injections were done once every 3 days in all groups; all the above mice were sacrificed on the 21st day, and the tumors were resected and weighed. Immunohistochemistry assay was performed to determine the CD31 expression in tumor tissues. One-way analysis of variance was used for comparisons among multiple groups, and least significant difference (LSD) - t test was used for multiple comparisons. Results:The proliferation rate of A431 cells significantly differed among the control group, 50 μmol/L resveratrol group, and 100 μmol/L resveratrol group ( F = 17.75, P = 0.017), and was significantly lower in the 50 μmol/L resveratrol group (66.53% ± 5.09%) and the 100 μmol/L resveratrol group (35.88% ± 4.28%) than in the control group (100%, LSD- t = 21.17, 29.04, P = 0.011, 0.004, respectively) ; the total length of vessel wall-like structures formed by A431 cells significantly differed among the 3 groups ( F = 21.37, P = 0.004), and was significantly lower in the 50 μmol/L resveratrol group (102.73 ± 11.36 μm) and the 100 μmol/L resveratrol group (37.83 ± 4.19 μm) than in the control group (185.26 ± 8.02 μm, both P < 0.05) ; the relative protein expression levels of ubiquitin, SUMO1, HIF-1α, and VEGFR also significantly differed among the 3 groups, the ubiquitin protein expression was significantly higher in the 50 μmol/L resveratrol group (2.09 ± 0.13) and the 100 μmol/L resveratrol group (3.53 ± 0.16) than in the control group (0.68 ± 0.11, both P < 0.05), while the protein expression of SUMO1, HIF-1α, and VEGFR was significantly lower in the 50 μmol/L resveratrol group (1.87 ± 0.13, 0.81 ± 0.06, 0.73 ± 0.09, respectively) and the 100 μmol/L resveratrol group (1.02 ± 0.11, 0.45 ± 0.06, 0.39 ± 0.05, respectively) than in the control group (3.10 ± 0.11, 0.97 ± 0.08, 0.98 ± 0.07, respectively, all P < 0.05). In the mice experiment, the weight of subcutaneous tumors and the proportion of CD31-positive cells in tumor tissues significantly differed among the control group, 1 mg/kg resveratrol group, and 2 mg/kg resveratrol group (weight: 3.29 ± 0.57 g, 2.91 ± 0.49 g, 2.55 ± 0.52 g; proportion: 76.24% ± 5.51%, 39.45% ± 5.48%, 12.07% ± 3.54%; F = 14.33, 15.34, P = 0.019, 0.021, respectively), and were significantly lower in the 1 mg/kg resveratrol group and 2 mg/kg resveratrol group than in the control group (all P < 0.05) . Conclusion:Resveratrol could inhibit tumor growth and neovascularization in tumor tissues, which were possibly associated with the inhibitory effect of resveratrol on neovascularization in cutaneous squamous cell carcinoma by suppressing the SUMOylation of HIF-1α protein via ubiquitin/ubiquitin-like protein modification pathways.

Objective To study and compare the oral microbiota and metabolites of children with Henoch Schonlein purpura(HSP)to identify specific microbiota and metabolites related to this disease and elucidate the pathogenesis of HSP.Methods Three groups of qualified subjects were included,including 20 in the HSP group,20 in the HSP nephritis(HSPN)group,and 20 in the control group.Perform high-throughput 16S rRNA sequencing and metabolic profiling of saliva from each group to analyze the correlation between differential microbiota and differ-ential metabolites.Results(1)Compared with the control group,there was a significant difference in richness and diversity in the HSPN group(P＜0.05).At the same time,there was no significant difference in richness and diver-sity in the HSP group(P＞0.05).Compared with the HSP group,the abundance,and diversity of the HSPN group were significantly increased(P＜0.05).At the genus level,the proportion of Streptococcus in each group is the high-est.Compared with the control group,there was no significant correlation between the HSP group and the genus of bacteria.In contrast,the HSPN group showed a significant increase in the genera of Pseudomonas and Parabacteroi-des(P＜0.05).Compared with the HSP group,the abundance of Pseudomonas and Parabacteroides in the HSPN group was significantly increased(P＜0.05).(2)Compared with the control group,the HSPN group had 12 differen-tial metabolites involving nine metabolic pathways,such as phenylalanine metabolism;There was no significant dif-ference in metabolites and no metabolic pathway in the HSP group.Compared with the HSP group,the HSPN group has 15 differential metabolites involving nine metabolic pathways,such as phenylalanine metabolism.(3)In the HSPN and control groups,Pseudomonas and Parabacteroides negatively correlated with Phenylalanine metabolic pathway products.In the HSPN and HSP groups,Pseudomonas,Parabacteroides,and Phenylalanine metabolic path-way products were negatively correlated.The metabolites involved in phenylalanine metabolism in the oral cavity are 2-hydroxycinnamic acid,Phenylpyruvic acid,and N-acetyl-L-phenylalanine.Conclusion There is a significant dif-ference between HSPN and HSP children and healthy children.Streptococcus,Pseudomonas,and Parabacteroides may be one of the trigger factors of HSPN,and Phenylalanine metabolism may be one of the pathways in the patho-genesis of HSPN.Children with HSPN have a more pronounced imbalance in oral microbiota and greater differences in metabolic products than children with HSP.

Cardiac lipoma is rare and benign entities of heart, and often discovered after autopsy incidentally because most patients remain completely asymptomatic. The symptoms of cardiac lipoma depend on their location and size within the heart, such as dyspnea, chest pain, arrhythmia, and even sudden death. Surgical interventions usually have good results. Up till now, there has been no any relevant large-scale randomized controlled trial, and even no precise guideline for treatment. Surgical procedures often depend on patients' clinical manifestations and changes of hemodynamics in cardiac vessels in order to relief the symptoms as well as abort the progress of the disease. Therefore, early diagnosis and close follow-up are necessary for timely treatment. This article aims to summarize the imageological examinations for cardiac lipoma, including echocardiography, computed tomography, magnetic resonance imaging along with their characteristics and advantages, in order to get better clinical strategies.

Objective:To evaluate the short-term outcomes of aortic root repair using a pericardial autograft for acute type A aortic dissection.Methods:Between January 2020 and August 2021, 12 patients underwent aortic root repair using a pericardial autograft for type A aortic dissection. There were 8 males and 4 females, aged from 32.0 to 71.0 years, with a mean of (49.1±13.0) years, weight from 40.0 to 100.0 with a mean of (78.6±20.5) kg. Short-term outcomes were assessed by cardiopulmonary bypass time, cross-clamp time, circulatory arrest time, mechanical ventilation time, ICU time, postoperative stay time. The diameter of Valsalva sinus and aortic regurgitation were assessed before, after operation and during follow-up.Results:There were no postoperative and follow-up death. Cardiopulmonary bypass time was(256.4±60.6)min, ranging from 182.0 to 243.0 minutes; cross-clamp time was(195.0±52.5)minutes, ranging from 127.0 to 284.0 minutes; circulatory arrest time was(19.9±3.6)min, ranging from 15.0 to 25.0 min; mechanical ventilation time was (69.1±72.1)hours, ranging from 6.4 to 250.3 h; ICU time was (143.6±81.7) h, ranging from 56.9 to 288.0 h; postoperative stay time was (12.8±4.1) days, ranging from 8.0 to 20.0 days. One patient had transient paralysis (8.3%), and one patient needed hemofiltration due to acute kidney failure (8.3%), they all completely recovered before discharge. Follow-up time ranged from 10.0 to 21.0 months, with a median of 13 months. Heart function (NYHA) was I to II degrees. The mean diameter of aortic root was (36.7±5.8)mm(27.0-45.0 mm) preoperatively, (35.8±5.1)mm (25.0-44.8 mm) before discharge, and (35.7±5.9)mm (25.1-44.8 mm) during follow-up, respectively. There was no significant difference between them ( P>0.05). The preoperative aortic regurgitation was as follow: severe aortic regurgitation 2, moderate 1, mild to moderate 3, mild 1, trivial 1, none 4; postoperative aortic regurgitation: mild 2, trivial 2, none 8; follow-up aortic regurgitation: mild 3, trivial 1, none 8. Conclusion:Aortic root repair with a pericardial autograft is a safe and effective technique to treat acute type A dissection involving the sinus. Using this technique, residual dissection tissues could be significantly reduced, which subsequently decreases the risk of proximal bleeding, maintains the function of sinus, and increases long-term durability. Short-term results are satisfactory, and the long-term results need further follow-up.

Objective:To evaluate the short-term outcomes of cuff wrapping technique using remnant aortic wall in modified Bentall procedure.Methods:Between January 2018 and December 2018, 23 patients underwent modified Bentall procedure with the remnant aortic wall as a cuff to cover the sewing area of composite valved graft and the aortic annulus for proximal hemostasis. After the sewing ring of the composite valved graft was sutured to the aortic annulus by continuous suture, the remnant aortic wall was sutured to the graft just distal to the sewing ring by continuous suture. There were 21 males and 2 females, aged from 22 to 72 with a mean of(50.96±13.23)years. Short-term outcomes were assessed by cardiopulmonary bypass time, clamp aorta time, mechanical ventilation time, ICU time, postoperative stay time, effusion drainage till the first postoperative day, Left ventricular end diastolic diameter(LVEDD), left ventricular ejection fraction(LVEF), and follow-up results.Results:There were no postoperative and follow-up death. Cardiopulmonary bypass time was(157.74±39.85)min, ranged from 114 to 275 min; clamp aorta time(122.61±30.25)min, ranged from 84 to 212 min; mechanical ventilation time(11.65±3.08)h, ranged from 7.87 to 20.33 h; ICU time(81.43±45.88) h, ranged from 15.18 to 184.77 h; postoperative stay time(8.73±2.80)days, ranged from 6 to 15 days. Effusion drainage was(336.82±117.65)ml, ranged from 200 to 670 ml till the first postoperative day. Follow-up was performed from 19 to 30 months, with a mean of(23.17±3.17)months. There were significant differences between preoperative LVEDD and postoperative LVEDD before discharge[(49.78±6.21)mm vs.(58.78±10.54)mm, P<0.05]; There were a little decrease of follow-up LVEDD compared with postoperative LVEDD, but no significant difference between them[(48.87±4.63)mm vs.(49.78±6.21)mm, P>0.05] . There were a little decrease of postoperative LVEF compared with preoperative LVEF, but no significant difference between them(0.57±0.07 vs. 0.59±0.05, P>0.05). There were significant differences between follow-up LVEF and preoperative LVEF(0.62±0.04 vs. 0.57±0.07, P<0.05), postoperative LVEF before discharge( P<0.05). Conclusion:Cuff wrapping technique using remnant aortic wall in modified Bentall procedure has got good short-term results. This modification may be a simple, effective way in controlling proximal bleeding.

Objective:To evaluate the safety and effectiveness of adventitial inversion technique for root repair in patients with acute type A aortic dissection(ATAAD).Methods:Between 2015 and 2018, ATAAD patients with dissected root and underwent open surgery were included. The exclusion criteria were as follows: previous root intervention, traumatic dissection and patient underwent root replacement(Bentall or David procedure). 490 ATAAD patients were included, 366(74.69%) male and 124(25.31%) female, aged(51.28±10.99) years(range 24-77 years). The clinical data were retrospectively analyzed with ANOVA/ nonparametric test and Chi- square test. Follow-up mortality and reoperation were displayed with Kaplan- Meier curve. Results:All patients were technically divided into three groups: adventitial inversion(A), direct suture(B) and Cabrol-shunt(C). The mean age in group A was(53.05±11.09) years, whereas worse cardiac and renal function occurred in group C. The mean duration of HCA, CPB and ACC were shortest, with a highest average of minimum rectal temperature during surgical interval in group A. Postoperative complications and early mortality were similar among groups. There were no significant differences of mid-term mortality and reoperation among these three techniques. Though no late reintervention for aortic root was found in both group A and B, the root diameter was more stable in group A during follow-up period[(33.14±3.74)mm vs.(34.51±3.83)mm vs.(33.89±3.89)mm, P=0.008]. Conclusion:Adventitial inversion technique is safe and effective for root repair in patients with ATAAD, achieving satisfactory short- and mid-term effects.

Acute Stanford type A aortic dissection has the characteristics of acute onset, severe condition and high mortality. Once making a definite diagnosis, surgical treatment is needed as soon as possible. It is difficult for cardiac surgeons to treat the acute aortic dissection involving the aortic sinus, which is an important risk factor for death. Improving the surgical treatment for the aortic sinus can be a key to improving the prognosis. In this review, we will introduce the modified sandwich technique for acute Stanford type A aortic dissection and the prognosis, and summarize the experiences of different modified sandwich techniques. However, there is still no unified standardized technique in aortic root repair, and there is a lack of large studies with long-term follow-up, so it is necessary to further improve the aortic root repair techniques.