[Preparation of chitin/hyaluronic acid/collagen hydrogel loaded with mouse adipose-derived stem cells and its effects on wound healing of full-thickness skin defects in rats].

Objective: To prepare the chitin/hyaluronic acid/collagen hydrogel loaded with mouse adipose-derived stem cells and to explore its effects on wound healing of full-thickness skin defects in rats. Methods: The research was an experimental research. Chitin nanofibers were prepared by acid hydrolysis and alkaline extraction method, and then mixed with hyaluronic acid and collagen to prepare chitin/hyaluronic acid/collagen hydrogels (hereinafter referred to as hydrogels). Besides, the hydrogels loaded with mouse adipose-derived stem cells were prepared. Thirty male 12-week-old guinea pigs were divided into negative control group, positive control group, and hydrogel group according to the random number table, with 10 guinea pigs in each group. Ethanol, 4-aminobenzoic acid ethyl ester, or the aforementioned prepared hydrogels without cells were topically applied on both sides of back of guinea pigs respectively for induced contact and stimulated contact, and skin edema and erythema formation were observed at 24 and 48 h after stimulated contact. Adipose-derived stem cells from mice were divided into normal control group cultured routinely and hydrogel group cultured with the aforementioned prepared hydrogels without cells. After 3 d of culture, protein expressions of platelet-derived growth factor-D (PDGF-D), insulin-like growth factor-Ⅰ (IGF-Ⅰ), and transforming growth factor ß1 (TGF-ß1) were detected by Western blotting (n=3). Eight male 8-week-old Sprague-Dawley rats were taken and a circular full-thickness skin defect wound was created on each side of the back. The wounds were divided into blank control group without any treatment and hydrogel group with the aforementioned prepared hydrogels loaded with adipose-derived stem cells applied. Wound healing was observed at 0 (immediately), 2, 4, 8, and 10 d after injury, and the wound healing rate was calculated at 2, 4, 8, and 10 d after injury. Wound tissue samples at 10 d after injury were collected, the new tissue formation was observed by hematoxylin-eosin staining; the concentrations of interleukin-1α (IL-1α), IL-6, IL-4, and IL-10 were detected by enzyme-linked immunosorbent assay method; the expressions of CD16 and CD206 positive cells were observed by immunohistochemical staining and the percentages of positive cells were calculated. The sample numbers in animal experiment were all 8. Results: At 24 h after stimulated contact, no skin edema was observed in the three groups of guinea pigs, and only mild skin erythema was observed in 7 guinea pigs in positive control group. At 48 h after stimulated contact, skin erythema was observed in 8 guinea pigs and skin edema was observed in 4 guinea pigs in positive control group, while no obvious skin erythema or edema was observed in guinea pigs in the other two groups. After 3 d of culture, the protein expression levels of PDGF-D, IGF-I, and TGF-ß1 in adipose-derived stem cells in hydrogel group were significantly higher than those in normal control group (with t values of 12.91, 11.83, and 7.92, respectively, P<0.05). From 0 to 10 d after injury, the wound areas in both groups gradually decreased, and the wounds in hydrogel group were almost completely healed at 10 d after injury. At 4, 8, and 10 d after injury, the wound healing rates in hydrogel group were (38±4)%, (54±5)%, and (69±6)%, respectively, which were significantly higher than (21±6)%, (29±7)%, and (31±7)% in blank control group (with t values of 3.82, 3.97, and 4.05, respectively, Pvalues all <0.05). At 10 d after injury, compared with those in blank control group, the epidermis in wound in hydrogel group was more intact, and there were increases in hair follicles, blood vessels, and other skin appendages. At 10 d after injury, the concentrations of IL-1α and IL-6 in wound tissue in hydrogel group were significantly lower than those in blank control group (with tvalues of 8.21 and 7.99, respectively, P<0.05), while the concentrations of IL-4 and IL-10 were significantly higher than those in blank control group (with tvalues of 6.57 and 9.03, respectively, P<0.05). The percentage of CD16 positive cells in wound tissue in hydrogel group was significantly lower than that in blank control group (t=8.02, P<0.05), while the percentage of CD206 positive cells was significantly higher than that in blank control group (t=7.21, P<0.05). Conclusions: The hydrogel loaded with mouse adipose-derived stem cells is non-allergenic, can promote the secretion of growth factors in adipose-derived stem cells, promote the polarization of macrophages to M2 phenotype in wound tissue in rats with full-thickness skin defects, and alleviate inflammatory reaction, thereby promoting wound healing.

[Clinical characteristics and diagnosis and treatment strategies of patients with severe traumatic aortic injury].

Objective: To investigate the clinical characteristics and diagnosis and treatment strategies of patients with severe traumatic aortic injury (TAI). Methods: A total of 25 patients with TAI, who hospitalized in our hospital between August 2005 to March 2021 and underwent thoracic aortic endovascular repair (TEVAR), were included in this retrospective study. According to the time from admission to TEVAR, the patients were divided into emergency TEVAR group (14 cases, TEVAR within 24 h of admission) and elective TEVAR group (11 cases, patients underwent surgery or fracture reduction and fixation first for serious injuries and then underwent TEVAR more than 24 h after admission). The general clinical data of patients, injury severity score (ISS), time from admission to intervention, total hospital stay, the proportion of closed chest drainage and the proportion of abdominal organ repair were obtained and compared. Clinical follow-up and 1-year postoperative aortic computed tomography angiography (CTA) were performed on the patients. Death, the occurrence of aortic adverse events and injury recovery were followed up and recorded. Results: The mean age of these 25 TAI patients was (41.4±14.4) years, 20 patients were males (80.0%). 21 patients (84.0%) had persistent chest and back pain, 17 (68.0%) had pleural effusion and 5 (20.0%) had mediastinal hematoma. The injury severity score (ISS) was significantly higher in the elective TEVAR group than in the emergency TEVAR group (24.9±14.4 vs. 35.5±9.3, P=0.044). The time from admission to intervention ((1.0±0.0) d vs. (3.4±0.9) d, P<0.001], the time from admission to TEVAR ((1.0±0.0) d vs. (11.5±13.8) d, P=0.030) and total hospital stay ((6.1±2.3) d vs. (26.8±7.7) d, P<0.001) were significantly longer in elective TEVAR group than in emergency TEVAR group. The proportion of thoracic closed drainage was significantly lower in the elective TEVAR group than in the emergency TEVAR group (9 (64.3%) vs. 2 (18.2%), P=0.042). The proportion of abdominal organ repair was significantly higher than in the emergency TEVAR group (0 vs. 4 (36.4%), P=0.026). All of 25 patients were discharged alive and followed up for (84.0±30.5) months. All patients survived and completed 1-year postoperation CTA. There were no aortic adverse events occurred, and no complications after surgery, and the fractures and organ injuries healed well. Conclusions: The clinical characteristics of severe TAI are acute multi-injuries combined with persistent chest and/or back pain, pleural effusion, and mediastinal hematoma. Timely diagnosis and treatment are important factors for the outcome. The treatment strategy for multi-injuries should give priority to dealing with life-threatening injuries. TEVAR is the primary treatment strategy for severe TAI and is related to satisfactory outcomes.

[Influencing factors analysis on live birth outcome of D3 cleavage stage frozen-thawed embryo after overnight culture and development of nomogram prediction model].

Objective: To analyze the factors affecting the live birth outcome of D3 cleavage stage frozen-thawed embryos after overnight culture, and establish a nomogram model to predict the live birth probability. Methods: The clinical data of assisted reproductive patients treated with D3 cleavage stage frozen-thawed embryo transfer in the First Affiliated Hospital of Zhengzhou University from January 2017 to July 2020 were analyzed retrospectively. A total of 5 456 patients were divided into modeling group and validation group according to the ratio of 7∶3. The modeling group [3 831 patients with average age of (33±6) years] was used to evaluate the independent risk factors of the patient's live birth outcome through multivariate logistic regression analysis and construct the nomogram prediction model. The validation group [1 625 patients with average age of (33±6) years] was used to verify and calibrate the performance of the model. Results: The results of multivariate logistic regression analysis showed that the risk factors related to live birth outcome of D3 frozen-thawed embryos after overnight culture included: female age (OR=0.901,95%CI:0.889-0.914,P<0.001), body mass index (BMI) (OR=0.979,95%CI:0.957-1.002,P=0.072), endometrial thickness on the transfer day (OR=1.121,95%CI:1.080-1.164,P<0.001), the number of transferred embryos (OR=2.192,95%CI:1.867-2.579,P<0.001) and embryo division resumed after overnight culture (OR=1.405,95%CI:1.213-1.627,P<0.001). The area under the curve (AUC) of the nomogram model in the modeling group was 0.716 and that in the validation group was 0.739.Both sets of calibration curves fited well with the ideal curve, which illustrated that the model had good predictive ability. Conclusions: The female age, BMI endometrial thickness on the transfer day, the number of transferred embryos and the embryo division resumed after overnight culture are risk factors for the live birth outcome of frozen-thawed embryos after overnight culture. The nomogram established based on the above factors can help predict the probability of live birth after frozen-thawed embryo transfer.

[Chronic kidney disease after adrenalectomy in a patient with primary aldosteronism].

We report one case of estimated glomerular filtration rate (eGFR) decline after taking unilateral adrenalectomy due to aldosterone adenoma. A 60-year-old male with 23-year history of hypertension was reported to the endocrinologist due to hypokalemia (serum potassium 3.01 mmol/L). Urine microalbumin/creatinine (ALB/CR) was 70.15 mg/g, serum creatinine was 82 µmol/L and eGFR was 89.79 mL/(min·1.73 m2). Random serum aldosterone was 172.2-203.5 ng/L, and random plasma rennin activity was 0-0.17 µg/(L·h). His captopril challenge test suggested that his aldosterone le-vels were suppressed by 8% (< 30%) and the adrenal enhanced computed tomography scan revealed a left adrenal tumor. The patient was diagnosed with primary hyperaldosteronism (PA), aldosterone adenoma and underwent left laparoscopic adrenalectomy. Histological examination confirmed adrenal cortical adenoma. One week after the operation, his serum creatinine was increased to 127 µmol/L compared with preoperative level; eGFR was 32.34 mL/(min·1.73 m2). His systolic blood pressure (SBP) was 110 mmHg and diastolic blood pressure (DBP) was 60 mmHg (hypotensive drugs discontinued), and serum potassium level was 5.22 mmol/L. At the end of the 2-year follow up, the serum creatinine of this patient remained at 109-158 µmol/L and eGFR fluctuated from 63.28-40.12 mL/(min·1.73 m2). PA is one of the most common causes of secondary hypertension. Several studies have reported renal function deterioration of PA patients after unilateral adrenalectomy, like the patient in this article. Age, preoperative plasma aldosterone concentration, albuminuria and preoperative potassium level might be significant predictors of a decrease in the eGFR. Growing evidence suggests that aldosterone could contribute to structural kidney damage, arterial injury and hemodynamic disorder. At the same time, patients with PA exhibit glomerular hyperfiltration and glomerular vascular hypertension, leading to the misinterpretation of renal function in PA patients as subtle kidney damage may be masked by the glomerular hyperfiltration before treatment. After a unilateral adrenalectomy, glomerular hyperfiltration by aldosterone excess is resolved and renal damage can be unmasked. In conclusion, kidney function deterioration after adrenalectomy can be detected in some patients with PA. Thus, accurate evaluation of kidney function in patients with PA may be essential, especially for those with preoperative risk factors for postoperative renal impairment. After unilateral adrenalectomy, close monitoring of renal function and adequate management are required for PA patients.

Synergistic effects ofα-Fe2O3nanoparticles and Fe-doping on gas-sensing performance of NiO nanowires and interface mechanism.

High surface area nickel oxide nanowires (NiO NWs), Fe-doped NiO NWs andα-Fe2O3/Fe-doped NiO NWs were synthesized with nanocasting pathway, and then the morphology, microstructure and components of all samples were characterized with XRD, TEM, EDS, UV-vis spectra and nitrogen adsorption-desorption isotherms. Owing to the uniform mesoporous template, all samples with the same diameter exhibit the similar mesoporous-structures. The loadedα-Fe2O3nanoparticles should exist in mesoporous channels between Fe-doped NiO NWs to form heterogeneous contact at the interface of n-typeα-Fe2O3nanoparticles and p-type NiO NWs. The gas-sensing results indicate that Fe-dopant andα-Fe2O3-loading both improve the gas-sensing performance of NiO NWs sensors.α-Fe2O3/Fe-doped NiO NWs sensors presented the highest response to 100 ppm ethanol gas (55.264) compared with Fe-doped NiO NWs (24.617) and NiO NWs sensors (3.189). The donor Fe-dopant increases the ground state resistance and the absorbed oxygen content in air.α-Fe2O3nanoparticles in electron depletion region result in the increasing resistance in ethanol gas and decreasing resistance in air. In this way,α-Fe2O3/Fe-doped NiO NWs sensor presents the excellent gas-sensing performance due to the formation of heterogeneous contact at the interface.

[Clinical outcomes of intracytoplasmic sperm insemination with micro amount frozen-thawed diagnostic sperm in the treatment of azoospermia].

Objective: To investigate the efficacy and clinical outcomes of intracytoplasmic sperm injection (ICSI) with micro amount frozen-thawed diagnostic sperm obtained by microdissection testicular sperm extraction (microTESE), percutaneous epididymal sperm as-piration (PESA) and testicularsperm extraction (TESA) in the treatment of azoospermia. Methods: A retrospective analysis was performed on 736 ICSI cycles of azoospermia patients.In Reprocluctive Medicine Center of the First Affiliated Hospital of Zhengzhou University from January 2018 to December 2019. Including 199 ICSI cycles (microTESE 47cycles, PESA 75cycles and TESA 77 cycles) with micro amount frozen-thawed diagnostic sperm and 537 ICSI cycles (microTESE 23 cycles, PESA 111 cycles and TESA 403 cycles) with fresh micro amount sperm. The general conditions, embryo development conditions and clinical outcomes of patients were compared between and within the two groups. Results: The recovery rate of PESA group was significantly lower than that of TESA group (89.3% vs 98.7%), P<0.05. The rate of 2PN in the fresh control group was significantly higher than that in the experiment group (75.5% vs 71.3%) and the rate of 2PN in the fresh microTESE and PESA groups were also significantly higher than those of the frozen-thawed microTESE and PESA groups (74.2% vs 64.6%) and (78.5% vs 72.4%), P<0.05. Both the rate of D5 blastocyst formation and high quality blastocyst in the fresh group were significantly lower than that in the experiment group (26.9% vs 32.9%) and (15.1% vs 18.0%), P<0.05; both the rate of early cleavage and blastocyst formation in the fresh microTESE group were significantly lower than that in the frozen-thawed microTESE group (55.1% vs 68.3%) (27.3% vs 39.3%), P<0.05. Both the rate of 8 cells embryos and blastocyst formation in the fresh TESA group were significantly lower than those of the TESA frozen-thawed group (41.3% vs 46.0%) (26.5% vs 32.4%), P<0.05. There was no significant difference in pregnancy rate and planting rate between or within the groups(P>0.05). The abortion rate in the frozen-thawed group was significantly higher than the fresh group (12.0% vs 4.0%), P<0.05, especially the abortion rate in the PESA frozen-thawed group was significantly higher than the fresh group (18.0% vs 1.7%), P<0.05. There was no significant difference in gender, weight and body length between the fresh group and the frozen-thawed group (P>0.05), but there were two malformed babies born in the frozen-thawed group. Conclusions: Frozen-thawed microinjection of diagnostic microspermatozoa is a feasible method for the treatment of asthenospermia.There was on significonty difference in pregnancy rate and planting rate between of with in the groups. However, significantly higher than the fresh PESA group of the influence on offspring needs to be further studied.

[Effects of different ages and body mass index on embryo development time kinetic parameters and embryo development potential].

Objective: To investigate the effects of age and body mass index (BMI) on embryo development time kinetic parameters, embryo development potential and clinical pregnancy outcomes. Methods: A retrospective study was conducted to analyze the data of 6 294 embryos from 832 patients who underwent in vitro fertilization and embryo transfer (IVF-ET) in the Reproductive Medicine Center of the First Affiliated Hospital of Zhengzhou University from September 2016 to November 2018. According to the age, they were divided into two groups:<35-year-old group (655 cases, 5 076 embryos), ≥35-year-old group (177 cases, 1 218 embryos). According to the BMI, they were divided into three groups: low body mass group (BMI<18.5 kg/m(2), 47 cases, 355 embryos), normal body recombination (18.5-23.9 kg/m(2), 517 cases, 3 813 embryos), hyperrecombination (BMI>23.9 kg/m(2), 268 cases, 2 126 embryos). Embryo development time kinetic parameters, embryo development potential and clinical pregnancy outcomes in each group were compared. Results: Embryo development to 3 cells, 4 cells were faster in <35-year-old group than in ≥35-year-old group. The blastocyst formation rate, high-quality blastocyst formation rate, pregnancy rate, implantation rate, delivery rate, live birth rate, and abortion rate were all statistically significant (all P<0.05). There were no significant differences in normal fertilization rate, cleavage rate, embryo utilization rate, high quality embryo rate, pregnancy rate, implantation rate, abortion rate, delivery rate, live birth rate between the three BMI groups (all P>0.05). Conclusions: The age has an effect on the partial embryo development time kinetic parameters, but BMI has a little effect on it.

[Prediction of clinical outcomes of patients with surplus cleavage-stage embryos for blastocyst culture after D3 embryo transfer].

Objective: To find the best strategy of embryo transfer, so as to provide theoretical basis for improving the clinical outcomes of in vitro fertilization-Embryo transfer (IVF-ET), we investigate the blastocyst culture of surplus cleavage-stage embryos after D3 embryo transfer and the prediction of clinical outcomes with or without blastocyst formation. Methods: A retrospective study was conducted on 3 568 patients who underwent IVF-ET in the Reproductive Medicine Center of the First Affiliated Hospital of Zhengzhou University from January 2016 to May 2018, whotransplanted two embryos in D3 with blastocyst culture of surplus cleavage-stage embryos, according to their age, they were divided into three groups: <35 years old group, 35-38 years old group, and>38 years old group.And according to the presence or absence of blastocyst formation, they were also divided into two subgroups: blastocyst formation group and non-blastocyst formation group. The embryo development and clinical outcomes in each group were compared. Results: (1) Comparisons of the embryo development in the three age groups with the first cycle. The total fertilization rate, cleavage rate and high quality embryo rate of the blastocyst formation group in the three groups were higher than those in the non-blastocyst formation group, P<0.05; In<35 years old group, the embryo utilization rate (75.0% vs 70.6%), pregnancy rate (74.9% vs 70.3%), planting rate (53.6% vs 48.6%), delivery rate (66.7% vs 61.1%) and live birth rate (66.5% vs 61.0%) of the blastocyst formation group were higher than those in the non-blastocyst formation group, P<0.05. (2) Comparisons of embryo development in the three age groups with multiple cycles (≥2 cycles). In<35 years old group, the total fertilization rate (75.0% vs 70.6%),delivery rate (62.7% vs 43.8%) and live birth rate (62.7% vs 43.8%) of the blastocyst formation group were significantly higher than those in the non-blastocyst formation group, P<0.05; In>38 years old group, the pregnancy rate (56.3% vs 25.8%), implantation rate (34.4% vs 14.5%), delivery rate (43.8% vs 11.3%), live birth rate (43.8% vs 11.3%) of the blastocyst formation group were higher than those in the non-blastocyst formation group, P<0.05. Conclusions: The results of blastocyst culture in different groups can predict the outcomes of embryo transfer in D3. For patients<35 years old with the first cycle, the clinical outcomes of the blastocyst formation group after D3 embryo transfer is better than that of the non-blastocyst formation group. For Patients with multiple cycles (≥2 cycles),the clinical outcomes of the embryo formation group is superior to that of the non-blastocyst formation group<35 years old or>38 years old.

Effects of early-cleavage embryo transfer on in vitro fertilization-embryo transfer pregnancy outcomes.

PURPOSE: To observe the effects of early-cleavage embryo transfer (ET) on pregnancy outcomes in vitro fertilization-embryo transfer (IVF-ET). MATERIALS AND METHODS: The data of 6,548 two pro-nucleate (2PN) embryos and 968 patients who underwent IVF or intracytoplasmic sperm injection (ICSI) were analyzed. Of the 968 cycles, early-cleavage embryos were used in 432 cycles (early-cleavage group), late-cleavage embryos were used in 246 cycles (late-cleavage group), and both early and late-cleavage embryos were used in 290 cycles (mixed group). RESULTS: High-quality embryo rate was significantly higher in early-cleavage group than in late-cleavage group (82.74% vs 59.83%; p < 0.01). Both clinical pregnancy and implantation rates in IVF or ICSI were significantly higher in early-cleavage group than in late-cleavage group (all p < 0.01). In ICSI, both clinical pregnancy and implantation rates were significantly higher in mixed group than in late-cleavage group (all p < 0.05). CONCLUSION: Early-cleavage ET can improve pregnancy outcomes in IVF or ICSI.