[Objective]This study aimed to improve the existing semen processing methods in the field of reproductive male medicine,particularly focusing on the 300×g 20 min treatment condition in the double-layer density gradient method,to enhance fertilization outcome.[Methods]Semen specimens from 1623 patients undergoing assisted reproductive techniques at the Reproductive Medicine Center of the Sixth Affiliated Hospital of Sun Yat-sen University from July and September 2020 and March and May 2022 were collected for preliminary experiments.Four different double-layer density gradient methods(200×g 10 min,200×g 20 min,300×g 10 min,and 300×g 20 min)were compared for sperm DNA fragmentation rates and recovery rates after processing.Subsequently,the optimal method was selected as the new approach and compared with the current method in use(300×g 20 min double-layer gradient method)to assess any statistical differences in fertilization rates.Further optimization to a single-layer density gradient method was performed based on the new method and compared with the double-layer density gradient method to determine any statistical differences.Experimental conditions were strictly controlled for temperature,centrifugation speed,and duration,with the quantity and processing conditions of each sample recorded.[Results]Among the four double-layer density gradient methods,the sperm DNA fragmentation rate was lower with the 300×g 10 min treatment compared to 300×g 20 min while ensuring sufficient sperm recovery rates.Consequently,the 300×g 10 min method was selected as the new approach for experimentation.Results indicated that the total fertilization rate and 2 pronuclei(2PN)fertilization rate with the new 300×g 10 min method were higher than with the 300×g 20 min method,the difference was statistically significant(P<0.05).Although the cleavage rate with 300×g 10 min was slightly higher than 300×g 20 min,the difference was not statistically significant(P>0.05).The total fertilization rate and 2PN fertilization rate were slightly higher with the single-layer density gradient method compared to the double-layer density gradient method,but the difference was not statistically significant(P>0.05).The cleavage rate with the single-layer density gradient method was higher than the double-layer density gradient method,and the blastocyst formation rate is lower than that of the double-layer density gradient method,and the differences are statistically significant(P<0.05).[Conclusion]The 300×g 10 min double-layer density gradient method successfully improved total fertilization rates,2PN fertilization rates,and cleavage rates compared to the existing 300×g 20 min method,while reducing the time required for semen optimization processing.Although the single-layer density gradient method improves the cleavage rate,and saves reagent costs and operation time,its blastocyst formation rate has decreased.These findings provide valuable guidance and insights for semen processing methods in the field of reproductive andrology.

Objective To compare the clinical characteristics of cirrhosis with or without portal vein thrombosis(PVT),and to analyze the therapeutic effect of transjugular intrahepatic portosystemic shunt(TIPS)in treating cirrhosis with or without PVT.Methods The clinical data of 193 patients with cirrhosis complicated by gastrointestinal bleeding,who received TIPS from October 2018 to October 2022,were retrospectively analyzed.According to the presence or absence of PVT before TIPS,the patients were divided into non-PVT group(n=118)and PVT group(n=75).After TIPS,the patients were followed up at one,3,6 months and every 6 months thereafter.The effect of PVT on the clinical characteristics of cirrhosis patients and on the therapeutic efficacy after TIPS were analyzed.Results The success rate of TIPS was 100％in both groups.The proportion of carrying out splenectomy or partial splenic artery embolization(PSE)in PVT group was 26.7％(20/75),which was obviously higher than 13.6％(16/118)in non-PVT group,the difference between the two groups was statistically significant(x2=5.192,P=0.023).In PVT group the preoperative Child-Pugh score,the model of end-stage liver disease(MELD)score and serum sodium model of end-stage liver disease(MELD-Na+)score were(8.1±1.9)points,(9.2±8.0)pointsand(9.2±8.0)points respectively,which in non-PVT group were(7.4±1.9)points,(7.7±5.8)points and(7.7±5.8)points respectively,the differences between the two groups were statistically significant(all P＜0.05).The incidence of overt hepatic encephalopathy in PVT group was 33.3％(25/75),which was strikingly higher than 19.5％(23/118)in non-PVT group,the difference between the two groups was statistically significant(P=0.030).No statistically significant differences in postoperative survival rate,rebleeding rate and stent dysfunction rate existed between the two groups(all P＞0.05).Conclusion For the treatment of cirrhotic patients with PVT complicated by gastrointestinal bleeding,TIPS is clinically safe and effective.In cirrhotic patients with PVT,the worse the liver function is,the higher the incidence of overt hepatic encephalopathy after TIPS will be.

Objective To compare and analyze the clinical characteristics and efficacy of transjugular intrahepatic portosystemic shunt(TIPS)in the treatment of liver cirrhosis with different portal vein thrombosis(PVT)grades.Methods A retrospective analysis was performed on 75 patients with liver cirrhosis and gastrointestinal bleeding who received TIPS.According to the Yerdel scale of PVT,the patients were divided into type Ⅰ(34 cases),type Ⅱ(25 cases)and type Ⅲ(16 cases).The patients were followed up 1,3,6 months after TIPS and every 6 months thereafter to compare the clinical data and the efficacy of TIPS in three types of PVT patients.Results The success rate of TIPS in three types of patients was 100％.There were differences in platelet to lymphocyte ratio(PLR)and proportion of different Child-Pugh grades among the three types of patients(P＜0.05).After TIPS,portal vein pressure was decreased compared with that before TIPS(P＜0.001).However,there were no significant differences in postoperative survival rate,rebleeding rate,over hepatic encephalopathy rate,stent dysfunction rate,thrombus complete recanalization rate and thrombus recurrence rate(P＞0.05).Conclusion The success rate of TIPS in three types of patients is higher,and the portal vein pressure is decreased significantly after TIPS,but there are no significant differences in the postoperative efficacy.Although the implementation of TIPS in cirrhotic PVT patients is challenging,it is still worth the effort to reshape the portal vein for the benefit of patients.

Objective To investigate the effect of altered oxidative stress system on liver function after partial splenic embolization(PSE).Methods Twenty-nine patients with liver cirrhosis and hypersplenism who received PSE were selected.Peripheral venous blood was drawn from the patients before and at 1 week,1 month,and 3 months after PSE,and the indexes of oxidative stress system factors including malondialdehyde(MDA),superoxide dismutase(SOD),advanced oxidiation protein products(AOPPs),and gluta-thione peroxidase(GSH-Px)were detected,as well as liver function indexes.Results There were positive correlation between SOD activity and total bilirubin(TBil)and model for end-stage liver disease(MELD)scores at 1 week postoperatively(TBil:r=0.725,P＜0.05;MELD:r=0.764,P＜0.05).There was positive correlation between GSH-Px activity and alanine aminotransferase(ALT)at 1 month postoperatively(r=0.777,P＜0.05),however,the AOPPs was negatively correlated with ALT and aspartate aminotransferase(AST)at 3 months postoperatively(ALT:r--0.900,P＜0.05;AST:r=-0.957,P＜0.05).Conclusion PSE can improve the body oxidative stress system and enhance the body's antioxidant response,and then improve the liver function.

Objective To observe the application effect of family-centered diversified extended services in treating retinopathy (ROP) of premature infants. Methods A total of 100 premature infants with ROP were selected and randomly divided into control group and observation group, with 50 cases in each group. The control group received routine nursing services, while the observation group received family-centered diversified extended services. The compliance of parents of infants, ocular parameters of infants, the 20-item Measure of Processes of Care (MPOC-20) score by parents and the Chinese version of Parenting Sense of Competence Scale (C-PSOC) score were compared between the two groups. Results The compliance of parents in the observation group was significantly better than that in the control group (P<0.05); one month after intervention, the decrease of intraocular pressure, the anterior chamber angle at 500 μm from the scleral spur (ACA500), central anterior chamber depth (CACD), and anterior chamber angle opening distance at 500 μm from the scleral spur (AOD500) in the observation group were significantly greater than those in the control group (P<0.05); after intervention, the MPOC-20 and C-PSOC scores of parents in the observation group were significantly higher than those in the control group (P<0.05). Conclusion Family-centered diversified extended services can improve the compliance of parents in participating in treatment and their satisfaction degree with nursing services, improve ocular conditions in ROP children, enhance parents′ sense of parenting competence, and have a positive effect on the prognostic recovery of ROP patients.

Objective:To investigate the expression of syndecan-1 (SDC1) in human venous malformations and the effect of syndecan-1 (SDC1) silences on migration, invasion and angiogenesis of human umbilical vein endothelial cells (HUVECs).Methods:Pathological samples of venous malformation were selected from 20 patients with hemangioma surgically resected in the Third Affiliated Hospital of Zhengzhou University from December 2020 to June 2021 (10 males and 10 females, aged 1-57 years, median age 7 years ). HUVECs was purchased from American Type Culture Collection (ATCC ). Immunohistochemistry was used to detect the expression of SDC1 in venous malformations. The cytological experiments of HUVECs cultured in vitro were divided into two groups, transfected SDC1 small interfering RNA (siRNA) (si-SDC1 group) and control siRNA (control group), respectively. The siRNA with the best silencing effect was selected by real-time quantitative PCR and Western Blot method for follow-up experiments. Then the ability of HUVECs migration, invasion and angiogenesis was detected by scratch test, Transwell migration and invasion test and angiogenesis test, respectively.Results:SDC1 was positive in 13 of the 20 venous malformations and was mainly expressed in the cytoplasm of venous endothelial cells. Compared with HUVECs cells in NC group, the expression of SDC1 mRNA and protein in HUVECs cells in si-SDC1 group decreased, indicated that the synthesized siRNA targeting SDC1 could silence the expression of SDC1 , and the ability of HUVECs migration, invasion and angiogenesis were enhanced in si-SDC1 group.Conclusions:SDC1 was positively expressed in venous malformations. After silencing SDC1, the migration, invasion and tube formation ability of HUVECs were enhanced.

Objective:To investigate the expression of syndecan-1 (SDC1) in human venous malformations and the effect of SDC1 silencing on the migration, invasion and angiogenesis abilities of human umbilical vein endothelial cells (HUVECs).Methods:Twenty surgically resected samples of venous malformation were collected from 20 patients with hemangioma at the Third Affiliated Hospital of Zhengzhou University from December 2020 to June 2021 (10 males and 10 females, aged 1-57 years, median age 7 years). HUVECs were purchased from American Type Culture Collection (ATCC). Immunohistochemistry staining was used to detect the expression of SDC1 in venous malformations. HUVECs cultured in vitro were divided into two groups, and were transfected with SDC1 small interfering RNA (siRNA) (si-SDC1 group) or control siRNA (control group), respectively. The siRNA with the best silencing effect was screened and tested by real-time quantitative PCR and Western blotting for follow-up experiments. The migration, invasion and angiogenesis abilities of HUVECs were measured by scratch test, Transwell migration and invasion test, and angiogenesis test, respectively. Results:(1) SDC1 was positive in 13(65.0%) of the 20 venous malformations and was mainly expressed in the cytoplasm of venous endothelial cells. (2) Compared with HUVECs in the control group, the expression of SDC1 mRNA and protein in HUVECs cells in si-SDC1 group both decreased ( P<0.05), indicating the synthesized siRNA targeting SDC1 could silence the expression of SDC1. (3) The migration, invasion and angiogenesis abilities of HUVECs were enhanced in si-SDC1 group( P<0.05). Conclusions:SDC1 was expressed in most venous malformations. After silencing SDC1, the migration, invasion and tube formation abilities of HUVECs were enhanced.

Objective:To investigate the expression of syndecan-1 (SDC1) in human venous malformations and the effect of syndecan-1 (SDC1) silences on migration, invasion and angiogenesis of human umbilical vein endothelial cells (HUVECs).Methods:Pathological samples of venous malformation were selected from 20 patients with hemangioma surgically resected in the Third Affiliated Hospital of Zhengzhou University from December 2020 to June 2021 (10 males and 10 females, aged 1-57 years, median age 7 years ). HUVECs was purchased from American Type Culture Collection (ATCC ). Immunohistochemistry was used to detect the expression of SDC1 in venous malformations. The cytological experiments of HUVECs cultured in vitro were divided into two groups, transfected SDC1 small interfering RNA (siRNA) (si-SDC1 group) and control siRNA (control group), respectively. The siRNA with the best silencing effect was selected by real-time quantitative PCR and Western Blot method for follow-up experiments. Then the ability of HUVECs migration, invasion and angiogenesis was detected by scratch test, Transwell migration and invasion test and angiogenesis test, respectively.Results:SDC1 was positive in 13 of the 20 venous malformations and was mainly expressed in the cytoplasm of venous endothelial cells. Compared with HUVECs cells in NC group, the expression of SDC1 mRNA and protein in HUVECs cells in si-SDC1 group decreased, indicated that the synthesized siRNA targeting SDC1 could silence the expression of SDC1 , and the ability of HUVECs migration, invasion and angiogenesis were enhanced in si-SDC1 group.Conclusions:SDC1 was positively expressed in venous malformations. After silencing SDC1, the migration, invasion and tube formation ability of HUVECs were enhanced.

Objective:To investigate the expression of syndecan-1 (SDC1) in human venous malformations and the effect of SDC1 silencing on the migration, invasion and angiogenesis abilities of human umbilical vein endothelial cells (HUVECs).Methods:Twenty surgically resected samples of venous malformation were collected from 20 patients with hemangioma at the Third Affiliated Hospital of Zhengzhou University from December 2020 to June 2021 (10 males and 10 females, aged 1-57 years, median age 7 years). HUVECs were purchased from American Type Culture Collection (ATCC). Immunohistochemistry staining was used to detect the expression of SDC1 in venous malformations. HUVECs cultured in vitro were divided into two groups, and were transfected with SDC1 small interfering RNA (siRNA) (si-SDC1 group) or control siRNA (control group), respectively. The siRNA with the best silencing effect was screened and tested by real-time quantitative PCR and Western blotting for follow-up experiments. The migration, invasion and angiogenesis abilities of HUVECs were measured by scratch test, Transwell migration and invasion test, and angiogenesis test, respectively. Results:(1) SDC1 was positive in 13(65.0%) of the 20 venous malformations and was mainly expressed in the cytoplasm of venous endothelial cells. (2) Compared with HUVECs in the control group, the expression of SDC1 mRNA and protein in HUVECs cells in si-SDC1 group both decreased ( P<0.05), indicating the synthesized siRNA targeting SDC1 could silence the expression of SDC1. (3) The migration, invasion and angiogenesis abilities of HUVECs were enhanced in si-SDC1 group( P<0.05). Conclusions:SDC1 was expressed in most venous malformations. After silencing SDC1, the migration, invasion and tube formation abilities of HUVECs were enhanced.

Objective:To evaluate the therapeutic effect of radiofrequency ablation in the treatment of severe hemangioma with thrombocytopenia and coagulation abnormality (Kasabach-Merritt phenomenon, KMP).Methods:From April 2018 to April 2021, 11 cases of KMP in our department were retrospectively analyzed. The patients underwent MRI, color ultrasound and hematology examination. Radiofrequency ablation was performed for all patients under general anesthesia. The comprehensive, multi-level and multi-point ablation was accurately implemented under the whole-process intraoperative monitoring of color ultrasound. Postoperative ice pad was applied for the treatment area, and dressing change was carried out every day.Results:All 11 cases of KMP were treated with radiofrequency ablation, including 5 males and 6 females, aged 1 d to 2 months, with a median age of 15 d. Intraoperative color Doppler ultrasound monitoring showed that the tumor was completely ablated in the single operation. All patients were followed up for 0.5 to 2 years. The tumor was cured without recurrence. The local function was unaffected. The local scar formation was mild and the appearance was satisfactory.Conclusions:Radiofrequency ablation has great clinical value in the treatment of KMP. It makes up the shortcomings and avoids the disadvantages of traditional surgery and other conservative treatment methods. Good clinical results can be achieved by radiofrequence ablation with satifactory function and appearance.