Objective To explore the effect and mechanism of penehyclidine hydrochloride (PHCD) on vascular endothelial injury in septic rats. Methods Fifty male SD rats were randomly divided into control group, lipopolysaccharide (LPS) induced sepsis group (model group), low dose PHCD (0.3 mg/kg) group, medium dose PHCD (1.0 mg/kg) group and high dose PHCD (3.0 mg/kg) groups, ten mice for each group. Normal saline was injected into the tail vein of the control group, and 10 mg/kg lipopolysaccharide (LPS) was injected into the tail vein of the rats in other groups to prepare the sepsis rat models. After the models were successfully established, low, medium and high doses (0.3, 1.0, 3.0 mg/kg) of PHCD solution were injected into the tail vein of the rats of corresponding groups. Wet/dry mass ratio (W/D) of lung tissue of rats in each group was measured, and ELISA was used to assay interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), IL-6 content and rat plasma angiopoietin 2 (Ang2) content in bronchoalveolar lavage fluid (BALF). HE staining was used to observe the pathological changes of lung tissues. Immunohistochemical staining was used to observe the expression of Ang2 in the right lung tissues. Western blot analysis was performed to detect Ang2 and vascular endothelial cadherin (VE-cadherin) protein in lung tissues. Results Compared with the control group, the W/D ratio of the lung tissues of rats in the model group and the contents of IL-1β, IL-6 and TNF-α in BALF were significantly increased; the lung tissues showed obvious pathological damage, with up-regulation of Ang2 expression and down-regulation of VE-Cadherin expression. Compared with the model group, the W/D ratio of the lung tissues of rats in three PHCD treatment groups and the contents of IL-1β, IL-6 and TNF-α in BALF were significantly reduced; the pathological damage of lung tissue was significantly reduced, with down-regulation of Ang2 expression and up-regulation of VE-cadherin expression. Conclusion PHCD can reduce LPS-induced lung inflammation in rats with sepsis by regulating the Ang2/VE-Cadherin pathway, thereby improving vascular endothelial injury.
Rats ; Mice ; Animals ; Male ; Lipopolysaccharides/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Angiopoietin-2/pharmacology* ; Interleukin-6/metabolism* ; Rats, Sprague-Dawley ; Lung ; Acute Lung Injury/metabolism* ; Sepsis/metabolism*

【Objective】 To analyze the influence of β-lactam antibiotics on RBC aging and clearance by detecting various indicators of aging and clearance on RBCs, as well as the differences in phagocytosis for erythrocytes before and after drugs treated in vitro. 【Methods】 RBCs were treated by β-lactam antibiotics, including Penicillin, Cefepime, Cefoperazone and Ceftazidime, and the changing of phosphatidylserine (PS) and clearance related CD markers, including CD35, CD47, CD55 and CD59 on the surface of the RBCs, were detected by flow cytometry at 0h and 24h after drugs treatment. The proportion of acanthocytes by microscope also at 0h and 24h after drugs treatment was calculated. The phagocytosis of drug-treated RBC was detected by monocyte monolayer assay (MMA). Untreated RBCs were incubated in PBS by the same condition as a negative control.The influence of β-lactam antibiotics on RBC aging and clearance by all the results above was studied. 【Results】 Compare to the untreated RBCs, the drug treated RBCs showed a higher PS level on the cell surface. The results showed by percentage as following(0 h vs 24 h): Penicillin 9.42% vs 93.30%, Cefepime 3.88% vs 57.27%, Cefoperazone 4.71% vs 75.75% and Ceftazidime 3.05% vs 43.19%. The acanthocytes ratio was as following(0 h vs 24 h): Penicillin 7.33% vs 86%, Cefepime 2.67% vs 52.67%, Cefoperazone 3.33% vs 67.67% and Ceftazidime 3.33% vs 90.67%. On the opposite, the clearance related CD markers, showed an obviously lower level after drugs treated(0 h vs 24 h): CD35: Penicillin 7.36% vs 11.87%, Cefepime 0.14% vs 28.51%, Cefoperazone 11.85% vs 21.55% and Ceftazidime 7.63% vs 8.73%; CD47: Penicillin 1.22% vs 9.13%, Cefepime 1.80% vs 0.86%, Cefoperazone 0.08% vs 6.85% and Ceftazidime 1.54% vs 5.50%; CD55: Penicillin 14.46% vs 44.31%, Cefepime 17.27% vs 38.41%, Cefoperazone 19.28% vs 33.28% and Ceftazidime 14.62% vs 34.13%; CD59: Penicillin 4.71% vs 20.56%, Cefepime 4.03% vs 7.60%, Cefoperazone 5.91% vs 22.38% and Ceftazidime 5.93% vs 30.89%. Drug-treated RBCs attached more to monocytes than untreated RBCs. 【Conclusion】 The β-lactam antibiotics could induce the changing of PS and the clearance of related CD markers on surface of RBCs. They also could lead acanthocytes and make the RBCs more susceptible to phagocytosis by monocytes. The β-lactam antibiotics could promote the RBCs aging and clearance, which might deteriorate the DIIHA.

【Objective】 To construct an in-vitro model of erythrocyte antibody-mediated complement activation, and establish quantitative detection methods based on flow cytometry and spectrophotometry, so as to explore the correlation of anti-body titers and complement activation speed, and provide a methodological basis for studying the adverse transfusion reactions of anti-body mediated complement hemolysis. 【Methods】 Mouse monoclonal antibody that recognized human C3b and fluorescent secondary antibody were used to label C3b fragments on erythrocytes, and the deposition of C3b fragments after complement activation was detected by flow cytometry. The absorbance at 540 nm of the supernatant in the complement activation reaction system was measured by spectrophotometry as the amount of hemoglobin released was related to the absorbance. 【Results】 The complement activation system was constructed according to the ratio of 3% red blood cell suspension (mixed for 6 people) 1∶anti-Tj^a 1∶complement 2. The repeatability was good (P value>0.05) as different red blood cell mixtures had been used to repeat the detection reaction system. When using 32×, 64× and 128× dilutions of anti-Tj^a mediated complement activation, the deposition of C3b fragments has been detected by flow cytometry at 30 s, 1 min and 2 min, respectively, and MFI peaked at 5 min, 10 min and 30 min, respectively. No obvious hemolysis has been observed within 1.5 h. 【Conclusion】 In vitro model of anti-Tj^a-mediated complement activation demonstrates the speed of complement activation is related to the concentration of antibody. At a certain antibody concentration, the speed of complement activation has been slowed down, and no obvious hemolysis observed.

【Objective】 To establish human hybridoma cell lines, secreting monoclonal antibody against antigens of Rh blood system, from a donor with rare D--phenotype. 【Methods】 Peripheral blood B lymphocytes of an O type female donor, lacking C/c/E/e antigens on her erythrocyte, were transformed with Epstein-Barr virus (EBVs). EBVs were harvested from the cultural supernatant of B95-8 cells. The transformed lymphoblastoid cell line (LCL) secreting antibodies to C antigens were picked up and then hybridized with the myeloma SHM-D33 using electric fusion technique. Hybridoma cells were selected by HATD-Ouabain(HOTD)(Hypoxantine, Aminopterin, Thymidine, 2-Deoxycytide, and Ouabain)culture medium, microplate antibody screening and limited dilution subcloning. The monoclonal antibody was assayed by serological test and was confirmed by flow cytometry (FCM). 【Results】 From the cultural supernatant of D--peripheral blood transformed B lymphocytes, 3A6-C6, which agglutinated with R1R1(DCe/DCe)O-type RBCs but not with R2R2(DcE/DcE)O-type RBCs, was screened and preliminarily identified as anti-C. We established a hybridoma cell line secreting anti-C immunoglobulin M from B cells of D--individual successfully after hybridization with SHM-D33 myeloma cells. 【Conclusion】 The study had laid the groundwork for future research and development of human monoclonal antibodies against Rh antigens of RBC in future for diagnosis and screening purpose.

Objective:To explore the clinical efficacy and safety of different surgical procedures of Mayo level Ⅳ inferior vena cava tumor thrombus(IVC-TT).Methods:The clinical and pathological data of 36 patients with Mayo level Ⅳ tumor thrombus were collected in three large clinical centers in China, including 18 cases in PLA General Hospital, 7 cases in Nanfang Hospital, and 11 cases in Renji Hospital. There were 25 males and 11 females.The median age was 56.5 years (53-67 years old). The average body mass index was 24.18±2.55 kg/m 2. The average diameter of renal tumors was 8.24±3.25 cm. The average length of inferior vena cava tumor thrombus was 12.89±2.50 cm. Mayo level Ⅳ tumor thrombus were divided into level Ⅳa and level Ⅳb (301 classification) based on the criterion of whether the proximal end of the thrombus has invaded the right atrium. Among them, level Ⅳa patients underwent robot-assisted inferior vena cava thrombectomy without cardiopulmonary bypass(CPB-free group, 6 cases). Level Ⅳb patients underwent robot-assisted inferior vena cava thrombectomy with cardiopulmonary bypass(CPB group, 12 cases) or cardiopulmonary bypass with deep hypothermic circulatory arrest assisted inferior vena cava thrombectomy(CPB/DHCA group, 18 cases). The baseline data of the three groups of patients were comparable. The perioperative results and long-term survival data after surgery were compared with different surgical methods for grade Ⅳcancer thrombosis. Results:All operations were successfully completed. Compared with the CPB group, the CPB-free group had a shorter first portal blocking time[17.5(15-36)min vs. 36.5(12-102)min, P=0.044], less intraoperative bleeding [2 350(1 000-3 000)ml vs. 3 500 (1 500-12 000)ml, P=0.043] and a lower allogeneic blood transfusion [1 250(500-2 000)ml vs. 2 185(700-5 800)ml, P=0.049]. Compared with the CPB/DHCA group, the CPB-free group had an advantage in reducing intraoperative allogeneic blood transfusion [1 250(500-2 000)ml vs. 2 700(1 200-10 000)ml, P=0.003]. There were no significant differences between groups in terms of duration of surgery and postoperative hospital stay. Among the 36 patients in this group, 23(64%) developed major complications (level Ⅲ or above), including 9 (25%) grade Ⅲ, 12 (33%) grade Ⅳ, and 2 (6%) grade Ⅴ. The CPB-free group had a relatively low complication rate of grade Ⅳ or above [ 17% (1/6) vs.42% (5/12) vs.44% (8/18)]. There were no statistical differences in median progression-free survival (16.4 vs.12.3 vs.18.0 months, P=0.695) and overall survival (30.1 vs.30.2 vs.37.7 months, P=0.674) between the groups. Conclusions:Robot-assisted inferior vena cava thrombectomy without cardiopulmonary bypass has the advantages of short ischemia time of organs, less intraoperative bleeding, and low incidence of major complications, which can be used as a safe and feasible surgical strategy for selected level Ⅳ tumor thrombus.

Labia minora resection is one of the key method to solve labia minora hypertrophy. And it is reported that wedge resection is becoming a research hotspot as a common method. By a comprehensive literature search, the purpose of this article is to study and analyze the improvement of wedge resection in the treatment of labia minora hypertrophy over the past few years. At the same time, the characteristics, complications, and postoperative outcomes of the different resection method will be discussed based on the previous data. It is concluded that wedge resection has achieved positive treatment effects in the therapy of labia minora hypertrophy.

Phalloplasty can be considered as a important way to reconstruct erectile function for female-to-male transsexuals, patients with penile defect or genital malformations. Meanwhile, penile prostheses, such as autologous tissue, artificial prosthesis and other tissues, play a key role in reconstructing erectile function. Thus, the purpose of this article is to analyze relevant research reports in recent years, since research goes deeper and improvements have been achieved in this field.

Objective:To explore the clinical effect of the composite reduction labiaplasty by divisional design.Methods:From January 2020 to February 2022, the clinical data of patients with complex labia minora and clitoris prepuce hypertrophy who were treated in the Medical Cosmetic Center of Beijing Friendship Hospital Affiliated to Capital Medical University were collected and analysed. The inner side of their labia minora showed no chalasis in the vertical direction, but the outer side were slack with folds, accompanied by clitoris prepuce hyperplasia. We adopted the composite reduction labiaplasty by divisional design. The outer side of the labia minora and the clitoral prepuce were considered as a whole, and a wedge-shaped incision line was designed to remove the excess tissue in the vertical direction. On this basis, an arc was projected on the inner side of the labia minora to form an arcuate incision line on the inner side. The composite reduction labiaplasty was performed with the combination of wedge resection and edge resection. The postoperative follow-up observations included: postoperative complications, the symmetry of the labia minora and clitoris prepuce, the improvement of frictional discomfort, the cleanliness of the labia minora and the clitoris prepuce, and the patient's satisfaction with the shape of the labia minora and clitoris prepuce. For patients with sexual experience, follow-up content also includes changes in sexual satisfaction.Results:A total of 17 female patients aged 20 to 35 years, with an average age of 27 years, were included in this study. Follow-up was conducted 2 months after the operation. All incisions healed by first intention with no sawtooth-like deformity at the incision margin, and had natural appearance. Patients felt that the bilateral labia minora and clitoral foreskin were completely symmetrical in 3 cases, basically symmetrical in 12 cases, and partially asymmetric in 2 cases. These 2 cases of partial asymmetry did not require further surgical repair. In all 17 patients, the frictional discomfort was completely resolved, and the labia minora and clitoral foreskin were easier to clean. The patients were very satisfied with the appearance in 5 cases, satisfied in 10 cases, and dissatisfied in 2 case. There were 15 patients with previous sexual life experience, of which 10 patients had improved sexual satisfaction, and 5 patients had no change in sexual satisfaction after the operation.Conclusions:Through the divisional design, wedge resection and edge resection are performed in the composite reduction labiaplasty, which can solve the friction and discomfort caused by the hyperplasia of labia minora and clitoris prepuce in a single operation, and achieve better aesthetic results at the same time.

Objective:To explore the clinical effect of the composite reduction labiaplasty by divisional design.Methods:From January 2020 to February 2022, the clinical data of patients with complex labia minora and clitoris prepuce hypertrophy who were treated in the Medical Cosmetic Center of Beijing Friendship Hospital Affiliated to Capital Medical University were collected and analysed. The inner side of their labia minora showed no chalasis in the vertical direction, but the outer side were slack with folds, accompanied by clitoris prepuce hyperplasia. We adopted the composite reduction labiaplasty by divisional design. The outer side of the labia minora and the clitoral prepuce were considered as a whole, and a wedge-shaped incision line was designed to remove the excess tissue in the vertical direction. On this basis, an arc was projected on the inner side of the labia minora to form an arcuate incision line on the inner side. The composite reduction labiaplasty was performed with the combination of wedge resection and edge resection. The postoperative follow-up observations included: postoperative complications, the symmetry of the labia minora and clitoris prepuce, the improvement of frictional discomfort, the cleanliness of the labia minora and the clitoris prepuce, and the patient's satisfaction with the shape of the labia minora and clitoris prepuce. For patients with sexual experience, follow-up content also includes changes in sexual satisfaction.Results:A total of 17 female patients aged 20 to 35 years, with an average age of 27 years, were included in this study. Follow-up was conducted 2 months after the operation. All incisions healed by first intention with no sawtooth-like deformity at the incision margin, and had natural appearance. Patients felt that the bilateral labia minora and clitoral foreskin were completely symmetrical in 3 cases, basically symmetrical in 12 cases, and partially asymmetric in 2 cases. These 2 cases of partial asymmetry did not require further surgical repair. In all 17 patients, the frictional discomfort was completely resolved, and the labia minora and clitoral foreskin were easier to clean. The patients were very satisfied with the appearance in 5 cases, satisfied in 10 cases, and dissatisfied in 2 case. There were 15 patients with previous sexual life experience, of which 10 patients had improved sexual satisfaction, and 5 patients had no change in sexual satisfaction after the operation.Conclusions:Through the divisional design, wedge resection and edge resection are performed in the composite reduction labiaplasty, which can solve the friction and discomfort caused by the hyperplasia of labia minora and clitoris prepuce in a single operation, and achieve better aesthetic results at the same time.

Phalloplasty can be considered as an important way to reconstruct erectile function for female-to-male transsexuals, patients with penile defect or genital malformations. Meanwhile, various types of penile supports such as autologous tissue, artificial prosthesis and other tissues, play a key role in reconstructing erectile function. The purpose of this article is to analyze relevant research reports in recent years, since research goes deeper and improvements have been achieved in this field.