Objective:To investigate the effect of tissue engineered bladder patch constructed by double-layer silk scaffold and adipose-derived stem cells (ADSCs) in the repair and reconstruction of bladder.Methods:This study was conducted from May 2020 to March 2021. The silk fibroin (SF) aqueous solution was obtained from silkworm cocoons, and a double-layer silk scaffold composed of silk fibroin film and silk fibroin sponge was further prepared. The rat ADSCs were isolated, cultured, and the ADSCs surface markers (CD29, CD90, CD45, CD106) were identified by flow cytometry. The ADSCs were planted on a double-layer silk scaffold to construct a tissue-engineered bladder patch. Thirty-six male SD rats were randomly divided into three groups: tissue engineered bladder patch group (SF-ADSCs group, n=15), double-layer silk scaffold group (SF group, n=15), control group ( n=6). The tissue engineered bladder patch (SF-ADSCs group) and double-layer silk scaffold (SF group) were wrapped on the omentum to promote vascularization. The vascularization was evaluated by HE and immunofluorescence staining. The wrapped tissue engineered bladder patch and double-layer silk scaffold were used to repair the defective bladder. In the control group (six rats), the incision was closed immediately after the bladder tissue fully exposed. At 4 weeks and 12 weeks after operation, the general morphology of bladder tissue and cystography were performed to evaluate the recovery of bladder morphology. After the graft was harvested, HE and Masson's trichrome staining and immunofluorescence staining were used to observe the regeneration of bladder wall tissue. Urodynamics was used to assess the recovery of bladder function at 12 weeks after operation. Results:The flow cytometry results confirmed that the isolated cells positively expressed CD29 and CD90, and there was no significant expression of CD45 and CD106. Gross observation and scanning electron microscope confirmed that the preparation of double-layer silk scaffold not only had a pore structure that was conducive to cell planting, but also had good toughness and was facilitated to surgical suture. The number (43.50±2.66) and area (0.73±0.03)% of vascular-like structures in the SF-ADSCs group after the omentum encapsulation was significantly higher than that in the SF group [(24.50±3.51), (0.55±0.05)%], and the difference was statistically significant ( P<0.05). At 4 weeks after bladder repair, the histological staining of the grafts in the SF-ADSCs and SF groups showed a large number of degraded fragments of double-layer silk scaffold. At 12 weeks, the morphology of the graft in the SF-ADSCs group showed uniform bladder morphology, which was similar to that of normal bladder tissue. Immunofluorescence staining showed that the continuous urothelial layer, abundant smooth muscle tissue, vascular structure and regenerated neurons could be observed in the SF-ADSCs group. Urodynamic test showed that the bladder maximum volume (0.74±0.03)ml and compliance (16.68±0.44)μl/cm H 2O in the SF-ADSCs group, which were better than that in the SF group [(0.47±0.05)ml, (14.89±0.37)μl/cm H 2O], but lower than that in the control group [(1.12±0.08)ml, (19.34±0.45)μl/cm H 2O], and the difference was statistically significant ( P<0.05). Conclusions:The tissue engineered bladder patch constructed with double-layer silk scaffolds and ADSCs could promote the morphological repair of bladder tissue, the regeneration of bladder wall structure and the recovery of bladder physiological function.

Objective:To prepare the whole bladder acellular matrix (BAM) using the self-designed perfusion decellularization system, and evaluate the feasibility of constructing the tissue engineering bladder with the adipose-derived stem cells (ADSCs).Methods:This study was conducted from October 2020 to April 2021. The self-designed perfusion decellularization system was used, and four different decellularization protocols (group A, group B, group C and group D) were formulated, according to the flow direction of the perfusate and the action time of different decellularization solutions. Among them, the urethral orifice of the bladder tissue was used as the outflow tract of the perfusion fluid in groups A and B. The top of the bladder was cut off and used as the outflow tract of the perfusion fluid in groups C and D. In groups A and C, 1% Triton X-100 was treated for 6 h, and 1% sodium dodecyl sulfate (SDS) was treated for 2 h. In groups B and D, 1% Triton X-100 was treated for 7 h, and 1% sodium dodecyl sulfate (SDS) was treated for 1 h. In addition, the tissue in the normal bladder group was directly obtained from the natural bladder tissue, which did not require perfusion, cryopreservation and thawing. The fast and efficient decellularization protocol was screened out through HE, DAPI, Masson trichrome and Alcian Blue staining and quantitative analyses to prepare the whole bladder scaffold. The prepared BAM was used as the scaffold material, and the ADSCs were used as the seeding cells to construct the tissue engineering bladder. HE and DAPI staining were used to observe the distribution of ADSCs on the BAM.Results:HE and DAPI staining showed that there was no obvious nuclear residue in the group C. Masson trichrome and Alcian Blue staining showed that the collagen structure and glycosaminoglycan were well preserved in the group C. There was no significant difference in bladder wall thickness between the group C and the normal bladder group [(975.44±158.62)μm vs.(1 064.49±168.52)μm, P > 0.05]. The DNA content in the group C [(43.59 ±4.59) ng/mg] was lower than that in the normal bladder group, group A, group B and group D [(532.50±26.69), (135.17±6.99), (182.49±13.69) and(84.00±4.38)ng/mg], and the difference was statistically significant ( P<0.05). The collagen content [(10.98 ± 0.29)μg/mg] and glycosaminoglycan content [(2.30±0.18)μg/mg] in group C were not significantly different with those in the normal bladder group [(11.69±0.49) and (2.36±0.09)μg/mg, P>0.05]. Scanning electron microscopy showed that a large number of pore structures could be observed on the surface of the prepared BAM in groups A-D and were facilitated to cell adhesion. The isolated and cultured ADSCs were identified by flow cytometry to confirm the positive expression of CD90 and CD29, and the negative expression of CD45 and CD106. Live/dead staining and CCK-8 detection confirmed that the prepared BAM in the group C had no cytotoxicity. HE and DAPI staining showed that a large number of ADSCs were distributed on the surface and inside of the tissue engineering bladder. Conclusions:The whole bladder shape BAM prepared by the self-designed perfusion decellularization system could be used as the scaffold material for bladder tissue engineering, and the constructed tissue engineering bladder could be used for bladder repair and reconstruction.

Objective To evaluate the clinical efficacy of portal venous thrombolysis by way of TIPS.Methods The clinical data of 40 patients with portal venous system thrombosis treated by TIPS at our department from May 2012 to May 2018 were retrospectively analyzed.There were 34 cases of via catheterdirected thrombolysis(7 cases by catheter-directed thrombolysis alone and 27 cases by way of TIPS before catheter-directed thrombolysis),and 6 cases via pharmaco mechanical thrombectomy (AngioJet);the postoperative complications of the two methods were followed up.Results The portal vein was opened in all 40 patients,and there were no major complications during the operation.One patient in the catheter-directed thrombolysis group developed acute liver failure after surgery.In the mechanical thrombolysis group,1 patient was discharged after small intestinal necrosis resection and intestinal fistula reconstruction.After 6-24 months of postoperative follow-up,6 patients in the group of thrombolysis suffered from shunt canal stricture.Conclusions It is a safe and minimally invasive method to treat portal venous system thrombosis through TIPS.Mechanical thrombolysis is more direct and rapid than catheter thrombolysis.

Objective:To analyze the clinical characteristics and treatment of patients with renovascular hypertension (RVH) caused by renal arterial fibromuscular dysplasia (FMD).Methods:Clinical data and treatment result of 38 patients with renal arterial FMD and RVH admitted to our hospital from Jan 2014 to Dec 2020 were reviewed.Results:A total of 38 patients were enrolled in this study. Renal artery CTA showed that 40 renal arteries were involved, among these 6 branches had multifocal stenosis, and 34 branches had focal stenosis. Thity-three patients received surgical treatment, of which 32 patients underwent percutaneous transluminal renal angioplasty (PTRA), and 1 patient with renal aneurysm underwent renal artery stent implantation combined with aneurysm coil embolization. Postoperative blood pressure was significantly lower than that before the operation [(129.79±17.63) mmHg vs. (178.52±28.63) mmHg, t=-11.42, P<0.001]. The mean follow-up time was 35.5 months. Renal artery restenosis occurred in 4 patients and underwent reintervention. Conclusion:For patients with renal arterial FMD and RVH, PTRA is safe and effective, especially for patients with focal lesions, with fair short and mid-term prognosis.

Objective:To evaluate the mid-term results of endovascular treatment for transplant renal artery stenosis (TRAS).Methods:The clinical and follow-up data of TRAS patients undergoing endovascular treatment at the First Affiliated Hospital of Zhengzhou University from Jan 2014 to Jan 2021 were retrospectively analyzed.Results:A total of 2 230 patients underwent kidney transplantation, 78 cases(3.6%) developed TRAS, among those 27 patients received endovascular treatment and followed-up from 12 to 80 months(mean 36 months). Thirteen patients (48.1%) underwent renal graft angiography and balloon dilatation, of which 2 patients underwent stent placement, 14 patients (51.9%) underwent renal graft angiography with balloon dilatation and stenting. The serum creatinine 2 weeks postoperatively and 12 months postoperatively were 127.6 μmol/L (47-220 μmol/L) and 103.4 μmol/L (63-166 μmol/L), respectively, significantly lower than the preoperative 217.1 μmol/L (98-541 μmol/L), ( P<0.05). Glomerular filtration rate (GFR) before surgery was 8.3-105.3 ml/min, 2 weeks and 12 months after surgery compared to 24.6-132.2 ml/min and 47.3-113.9 ml/min( P<0.05). The preoperative peak systolic velocity (PSV) of the transplanted renal artery during the systolic phase was 234 cm/s (75-457 cm/s), compared to 129 cm/s (52-290 cm/s) ( P<0.05) 2 weeks and 118 cm/s (57-300 cm/s) 12 months postoperatively ( P<0.05). During the follow-up period, 2 patients (7.4%) died of multiple organ failure. Conclusions:TRAS is the most common vascular complication after kidney transplantation. Endovascular treatment has a high success rate and low complication rate.

Objective: To analyze the clinical significance of contrast-enhanced ultrasound in the treatment of patients with carotid stenosis. Methods: The clinical data of 89 patients with carotid stenosis was retrospectively analyzed.The morphology and stenosis of carotid plaques were observed by contrast-enhanced ultrasound, and analyzing the relationship between the patient′s clinical symptoms and treatment options. Results: There were 66 males, 23 females, age ranging from 41 to 88 years.There were 147 plaques in 89 patients and 58 patients with bilateral lesions. The intensity of plaque ultrasound contrast was grade Ⅰ in 40 cases(27%), grade Ⅱ in 30(20%), grade Ⅲ in 31(21%), andgrade Ⅳ in 46(31%). The symptomatic group had higher CEUS strengths in grade Ⅲ(21.4%) and grade Ⅳ(37.9%). The difference was statistically significant between the two groups (P<0.05). Symptomatic group with high proportion of severe stenosis (44.7%) and occlusion (9.7%). It is narrower than that of asymptomatic group.The difference of stenosis was statistically significant between the two groups (P<0.05). Conclusion Contrast-enhanced ultrasound can dynamically and visually assess the distribution and density of carotid plaque morphology. It is useful for evaluating the treatment of patients with carotid stenosis.

Background Exposure to ambient nitrogen dioxide (NO₂) could increase the risks of small for gestational age (SGA) and large for gestational age (LGA). Nevertheless, previous published studies usually use a time period over relatively long durations as the exposure window, such as trimester-specific or gestational months, to identify adverse pregnancy outcomes related susceptible exposure windows for ambient air pollution. At present, no study has explored associations of weekly-specific ambient air NO₂ exposure around pregnancy with SGA and LGA. Objective To evaluate the associations of exposure to ambient NO₂ over the preconception and entire pregnancy period with risks of SGA and LGA, as well as to explore critical windows of NO₂ exposure by refining exposure period to specific weeks. Methods Based on a birth cohort established by the project Environmental and LifEstyle FActors iN metabolic health throughout life-course Trajectories (ELEFANT) situated in Tianjin, 10 916 singleton pregnant women whose dates of the last menstrual period and delivery were both between June 2014 and June 2016, and whose gestational age were within 24-42 completed gestational weeks were included in this study. Each pregnant woman's exposures to ambient NO₂ throughout 12 weeks before pregnancy and pregnancy period were matched with daily average NO₂ concentrations obtained from the Chinese air quality reanalysis datasets (CAQRA). Distributed lag models incorporated in Cox proportional hazard regression models were applied to explore the associations of maternal exposure to weekly ambient NO₂ throughout 12 weeks before pregnancy and pregnancy period with risks of SGA and LGA after controlling for potential confounders including maternal age, ethnicity, educational level, occupation, body mass index before pregnancy, residence, times of gravidity and parity, smoking, alcohol consumption, husband smoking, and season of conception. Hazard ratios (HRs) with 95% confidence intervals (CIs) were calculated per 3 μg·m⁻³ increase in ambient NO₂ concentrations. Results The average levels of maternal exposure to NO₂ over the preconception, first trimester, second trimester, third trimester, and entire pregnancy periods were (39.6±10.8), (42.7±10.5), (44.8±12.7), (37.7±11.1), and (41.6±4.8) μg·m⁻³, respectively. For a 3 μg·m⁻³ increase in NO₂ over the first trimester, the risk of SGA increased by 19.0% (95%CI: 8.0%-32.0%). For a 3 μg·m⁻³ increase in NO₂ over the preconception, first trimester, and entire pregnancy, the associated risks of LGA increased by 7.0% (95%CI: 1.0%-13.0%), 37.0% (95%CI: 29.0%-46.0%) and 19.0% (95%CI: 9.0%-31.0%), respectively. For SGA, the susceptible exposure windows for NO₂ were observed during the 7^th to 12^th preconceptional weeks and the 6^th to 12^th gestational weeks, with the strongest association found at the 12^th preconceptional week, when the risk of SGA increased by 6.0% (95%CI:3.2%-8.9%) for a 3 μg·m⁻³ increase in NO₂. For LGA, the susceptible exposure windows for NO₂ were observed during the 1^st to 12^th preconceptional weeks and the 1^st to 6^th gestational weeks, with the strongest association found at the 12^th preconceptional week, when the risk of LGA increased by 6.1% (95%CI: 4.5%-7.8%) for a 3 μg·m⁻³ increase in NO₂. Conclusion Exposure to ambient NO₂ is associated with increased risks of both SGA and LGA, and the most susceptible weekly exposure windows are nested within the 12 weeks before pregnancy and early pregnancy.