Aim To establish an allogenetic mouse skin trans-plant model,in order to provide a research model for immunosup-pressive drugs. Methods Skins from the ears of C57BL/6 mice were transplanted to the back of BALB/c mice and skin isografts ( BALB/c mice to BALB/c mice) were used as control. Cyclos-porin A( CsA) was used as a model compound to test the imm-nosuppresive effect on allogenetic graft rejection. Following the transplation and CsA treatment, the graft rejection score and graft skin survival rate were quantified. Four and nine days after transplantation,serum IL-4,IL-12 and IFN-γ levels were meas-ured using ELISA kits. Twelve days after transplantation, mice were sacrificed. The weight of spleen and thymus was obtained, and CD4 + and CD8 + population of spleenic T cells were ana-lyzed using flow cytometer. Histological features were assessed by hematoxylin-eosin( HE) staining of formalin-fixed, paraffin-em-bedded graft skins. Results After transplantion, the graft rejec-tion score increased and graft skin survival rate decreased gradu-allly. Serum IL-12 and IFN-γ levels of allograft mice increased markedly. Compared with those of isograft mice, mice with skin allograft displayed a significant increase in the percentage of the CD8 + T cell subpopulation. Remarkable inflammation, such as edema, inflammatory cell infiltration were observed in allograft mice. Compared with saline treated mice, CsA significantly re-duced the graft rejection score and improved survival rate of skin grafts. And also, CsA treated mice had smaller spleen and thy-mus. Mice that received high doses of CsA had significantly less CD8 + T cells than those treated with saline. Moreover, allograft skins in mice that received CsA had less inflammation. Conclu-sions Allogenetic mouse skin transplantation exhibits acute graft rejection. CsA can inhibit the rejection in a dose dependent manner.

Objective To investigate the values of urine neutrophil gelatinase associated lipocalin (NGAL), kidney injury molecular-1 (KIM-1) and interleukin-18 (IL-18) in the diagnosis of acute kidney injury (AKI) in children after cardiopulmonary by-pass (CPB). Methods Sixty-seven patients who had undergone CPB were recruited from March to June 2013 and assigned to acute kidney injury group (AKI group) or non-acute kidney injury group (non-AKI group) according to the pediatric RIFLE (pRIFLE) cri-teria. Serum and urine samples were collected from each patient at 30 min, 2 h, 4 h, 24 h, 48 h and 72 h after CPB for serum and urine creatinine, urine NGAL, KIM-1 and IL-18. All the data were evaluated by receiver operator characteristic curve (ROC) analysis and area under curve (AUC) analysis. Results Twenty-three cases (34.3%) had AKI in 67 children after CPB. Among them 15 cases were risk-stage AKI, 4 cases injury-stage AKI, 3 cases failure-stage AKI and 1 cases loss-stage AKI. The levels of urine NGAL/Ucr were higher in AKI group than those in non-AKI group at 4h, 48h and 72h after CPB (P<0.05). The cut-off value of NGAL/Ucr was 1.200 at 4 h after CPB, the sensitivity and specificity for prediction of AKI were 0.864 and 0.561, and the AUC was 0.671 (95%CI:0.537-0.804). The levels of urine KIM-1/Ucr were higher in AKI group than those in non-AKI group at 48h and 72 h after CPB (P<0.05). The cut-off value of KIM-1/Ucr was 1.162 at 24h after CPB, the sensitivity and specificity for prediction of AKI were 0.773 and 0.512, and the AUC was 0.698 (95%CI:0.563-0.834). The levels of IL-18/Ucr were higher in AKI group than those in non-AKI group at 4 h after CPB (P<0.05). The cut-off value of IL-18/Ucr was 0.04 at 4 h after CPB, the sensitivity and specificity for predici-ton of AKI were 0.773 and 0.561, and the AUC was 0.655 (95%CI:0.510-0.800). Conclusions It is indicated that urine NGAL, KIM-1 and IL-18 may have important clinical values for early prediction of AKI.

Objective To observe the expression levels of kidney injury molecule-1(KIM-1) in renal tissues of ischemia-reperfusion rats,and to explore the value in the diagnosis of acute kidney injury.Methods Rats were randomly divided into two groups,control(CON) group (n=64) and acute kidney ischemia reperfusion injury (AIKI) group (n=64).Rats were sacrificed following reperfusion 2h,6h,24h,48h,72h,1 week (w),2 w,and 4 w.The changes of morphology were checked on HE staining sections under light microscope.The extent of tubulointerstitial injury was determined by Sayhan classification.The distribution and expression of KIM-1 in renal tissue were observed by immunohistochemistry and Western blotting.Serum samples were collected and serum creatinine measurement was performed at different reperfusion time points.Results (1) Compared with the CON group,the renal tubulointerstitial injury scores of AIKI group were significantly higher at all times after reperfusion (P<0.01).(2) The expression of KIM-1 was consistent with the tubulointerstitial injury.The positive correlation between KIM-1 and the tubulointerstitial injury scores was significant(r=0.887,P=0.003).(3) Compared with the CON group,serum creatinine in AIKI group was significant higher at 2h,6h,24h,48h,72h after reperfusion (P<0.05).Serum creatinine had no correlation with the damage of renal tubulointerstitial.Conclusion The expression of KIM-1 increases significantly in renal ischemia reperfusion injury,and it is consistent with the tubulointerstitial injury.Compared with serum creatinine,KIM-1 may be a more accurate biomarker of renal damage.

Objective:To evaluate the detection of copy number variation (CNV) by chromosome microarray analysis (CMA) in fetuses with congenital anomalies of the kidney and urinary tract (CAKUT).Methods:A total of 1 929 fetuses who were ultrasonically found with CAKUT and underwent CMA from Guangdong Women and Children's Hospital and Health Institute were enrolled in this retrospective study from January 2016 to July 2020. These fetuses were divided into isolated CAKUT group ( n=1 567), CAKUT with soft markers group ( n=269), and CAKUT with other structural anomalies group ( n=93) for comparing the detection rate of pathogenic CNV using Chi-square test or Fisher exact test. Results:(1)The detection rate of all and pathogenic CNVs were 6.5%(125/1 929) and 4.8%(93/1 929), respectively. The total detection rate of CNV, clinically significant CNV and large chromosome structural variations in the CAKUT with other structural anomalies group were higher than those of the CAKUT with soft markers group and isolated CAKUT groups[31.2%(29/93), 11.5%(31/269) vs 4.2%(65/1 567), χ2=119.002; 18.3%(17/93), 9.0%(24/269) vs 3.6%(56/1 567), χ2=49.677; 9.7%(9/93), 2.2%(6/269) vs 0.3%(4/1 567), χ2=42.727; all P<0.001]. CAKUT with other structural anomalies group had a higher detection rate of pathogenic CNV (18.3%, 17/93) than the CAKUT with soft markers group (8.6%, 23/269) and the isolated CAKUT group [3.4%(53/1 567)] ( χ2=51.932, P<0.001). (2) The detection rate of pathogenic CNV was the highest in fetuses with enhanced renal echo (14.7%, 23/156), followed by renal enlargement (8.2%, 5/61), renal dysplasia (5.0%,13/261), polycystic renal dysplasia (5.0%, 13/261), and hydronephrosis (4.8%, 20/413). Fetuses with polycystic renal dysplasia, renal agenesis, fused kidney and hydronephrosis in the CAKUT with other structural anomalies group had a higher detection rate of pathogenic CNV than those in the isolated CAKUT group [3/9 vs 3.5%(8/230), 2/17 vs 1.3%(3/237), 1/8 vs 0.0%(0/59) and 3/18 vs 3.4%(12/344), all P<0.017]. The CAKUT with other structural anomalies group had a higher detection rate of pathogenic CNV than CAKUT with soft markers group in fetuses with enhanced renal echo [4/8 vs 12.8%(5/39), P<0.017]. (3) The top three microdeletion/microduplication syndrome were 17q12 microdeletion syndrome (36.6%, 34/93), 22q11.2 microdeletion syndrome (23.7%, 22/93), and 16p11.2 microdeletion syndrome (7.5%, 7/93) among those with pathogenic CNV. Conclusions:The risk of CNV in fetuses with isolated CAKUT, CAKUT with soft markers, and CAKUT with additional structural anomalies increased progressively. CMA might be a better choice in fetuses with hydronephrosis, enhanced renal echo, renal enlargement, renal hypoplasia, and multicystic renal dysplasia to improve the detection rate of CNV.

OBJECTIVETo investigate the new biomarkers of acute kidney injury, as well as to confirm the values of response gene to complement 32 (RGC-32) for early diagnosis of acute kidney injury by comparing the values of serum creatinine (Scr) and cystatin C (CysC) in children who had undergone cardiopulmonary bypass (CPB).

METHODSixty-seven patients who had accepted CPB were recruited from the cardiac surgery intensive care unit, Children's Hospital Affiliated to Shanghai Jiao Tong University from March to June 2013 and assigned to acute kidney injury group (group AKI) or non-acute kidney injury group (group non-AKI), on the basis of the definition by the pediatric RIFLE (pRIFLE) criteria. Also 30 healthy control children were recruited. Serum samples were taken regularly from each patient after CPB at 30 min, 2 h, 4 h, 24 h, 48 h and 72 h for RGC-32. Serum samples were tested by enzyme linked immunosorbent assay (ELISA) which was employed to determine the levels of serum RGC-32. Scr and CysC were analyzed by HITACHI 7180 automatic biochemical analyzer. All the data were analyzed by receiver operator characteristic curve (ROC) and area under curve (AUC).

RESULTThe incidence of AKI was 34% (23/67), including 15 cases with risk stage AKI, 4 cases with injury stage AKI, 3 cases with failure stage AKI, 1 cases with loss stage AKI. Three out of four subjects with Failure stage AKI and the one case with Loss stage all accepted renal replacement therapy. CPB group had a higher level of serum RGC-32 than that of pre-operation after CPB 30 minute [(2.88 ± 0.68) µg/L vs. (1.39 ± 0.31) µg/L, P < 0.05]. At the same time, comparing with the non-AKI group, the levels of serum RGC-32 were higher than that of controls 30 min, 2 h, 4 h, 24 h and 48 h after CPB (t = 2.560, 2.180, 2.818, 2.226, 3.017; P < 0.05). The values for the AUC were determined for RGC-32 as 0.770, 0.707, 0.768, 0.728,0.723 and 0.770 after CPB 30 min, 2 h, 4 h, 24 h, 48 h and 72 h. The values for sensitivity of serum RGC-32 30 min, 2 h and 4 h after CPB was 0.914, 0.824, 0.824 and the values for specificity of serum RGC-32 was 0.619, 0.667, 0.810, respectively. But the values for sensitivity of CysC was 0.625, 0.813, 0.813, and specificity 0.571, 0.619, 0.571, respectively. The values for sensitivity of Scr was 0.625, 0.625, 0.813 and specificity was 0.571, 0.571, 0.524, respectively.

CONCLUSIONThe sensitivity of serum RGC-32 for detecting AKI was much higher than that of Scr and serum CysC in children who had accepted CPB, and that RGC-32 may be a new biomarker for early detection of AKI. However, the conclusion needs to be further elucidated.

Acute Kidney Injury ; blood ; diagnosis ; etiology ; Area Under Curve ; Biomarkers ; blood ; Cardiopulmonary Bypass ; adverse effects ; Case-Control Studies ; Cell Cycle Proteins ; blood ; Creatinine ; blood ; Cystatin C ; blood ; Female ; Heart Defects, Congenital ; surgery ; Humans ; Infant ; Intensive Care Units, Pediatric ; Male ; Muscle Proteins ; blood ; Nerve Tissue Proteins ; blood ; Postoperative Complications ; Predictive Value of Tests ; Prospective Studies ; ROC Curve ; Sensitivity and Specificity

Objective:To explore the method and clinical efficacy of Flow-through latissimus dorsi musculocutaneous flap to repair children’s limb wounds.Methods:From January, 2017 to September, 2019, the Flow-through latissimus dorsi myocutaneous flap was used to repair the limb wounds in 9 children, which were 3 cases of upper extremity wounds and 6 cases of lower extremity wounds. The patients were 5-12 years old, with an average age of 9.4 years. The wound debridement was performed in the first stage, and the wound was covered with VSD. The second operation was performed 5-8 days later. Flow-through free latissimus dorsi muscle flap was used to repair the wound, and 6 patients combined with skin grafting. During the operation, both ends of the recipient area artery were trimmed into a downward slant. The proximal end of the recipient area blood vessel was coincided with the subscapular artery, the distal end was coincided with the circumflex scapular artery, and the thoracodorsal vein and the recipient area vein were coincided. One patient had 2 thoracodorsal veins, and 8 patients had 1 thoracodorsal vein. After the operation, the patients were given anti-infection, anticoagulation and antispasmodic treatment. And regular follow-up.Results:In this group, 8 flaps survived completely, and 1 had partial epidermal necrosis, which healed under the scab. Skin grafting survives well. Nine patients were followed-up for 3 to 28 months, 7 of which were followed-up in the outpatient clinic, and 2 by WeChat. CDU showed smooth blood flow in all patients 1 month after operation, in addition of 3 after 1 year. The flap had good blood flow and soft texture. Flap plastic surgery was performed in 3 cases.Conclusion:Using Flow-through latissimus dorsi musculocutaneous flap to repair the wounds of the children’s limbs, the flap survived well after the operation, and the blood circulation of the main vessel in the affected area was smooth, which had no obvious effect on the blood supply to the distal limb of the affected area.

Objective To understand the differences in the demand,preference,and tendency for elderly care services between urban and rural areas in the Pearl River Delta(PRD),and to provide reference for the planning and balanced allocation of elderly care resources in urban and rural areas.Methods Using the multi-stage stratified random sampling method,we selected 7 community health service centers in 2 prefecture-level cities in the PRD and conducted a questionnaire survey on the elderly care service demand,preference,and tendency among 1919 regular residents aged 60 years and above who attended the centers.Results A total of 641 urban elderly residents(33.4%)and 1278 rural elderly residents(66.6%)were surveyed in the PRD.The urban and rural elderly residents showed differences in the child number(χ2 =43.379,P<0.001),willingness to purchase socialized elderly care services(χ2 =104.141,P<0.001),and attitudes to the concept of raising child to avoid elderly hardship(χ2 =65.632,P<0.001).The proportion(71.8%)of rural elderly residents who prefer family-based elderly care was higher than that(57.1%)of urban elderly residents(χ2 =41.373,P<0.001).The proportion(62.2%)of urban elderly residents clearly expressing their willingness to choose institutions for elderly care was higher than that(44.0%)of rural elderly residents(χ2 =57.007,P<0.001).Compared with family-based elderly care,the willingness to choose institutional or community-based in-house elderly care was low among the urban elderly residents with surplus monthly household income or balanced income and expenditure;urban males,those with college education background or above,and those who purchased so-cialized elderly care services tended to prefer community-based in-house elderly care.In rural areas,the elderly residents who had local household registry were prone to choose institutional or community-based in-house elderly care,while those who had more than one child and those who were satisfied with the current living conditions were less willing to choose community-based in-house elderly care.Conclusions It is suggested that the urban-rural differences in the elderly care service demand,preference and tendency should be fully considered in the planning and allocation of urban and rural elderly care resources.Efforts remain to be made to develop diversified social elderly care services tailored to the characteristics of urban and rural areas.

Objective To predict the micro-level energy release rate in the rat femoral cortical bone and investigate the variation in the micro-level energy release rate with age.Methods Based on previous experimental data and numerical simulation of fracture modes for cortical bone,load-displacement curves and fracture modes measured by simulation and experiment were compared,and the micro-level energy release rates of rat femoral cortical bone at different months were predicted by back-calculation.Results It was predicted that the micro-level energy release rate of rat femoral cortical bone at 1-,3-,5-,7-,9-,11-,and 15-month age was 0.08-0.12,0.12-0.14,0.15-0.19,0.25-0.28,0.23-0.25,0.19-0.22,and 0.13-0.16 N/mm,respectively.Conclusions The decrease in the microlevel energy release rate with increasing age led to a decreasing failure load,indicating that the microlevel energy release rate is one of the main factors determining fracture occurrence;however,no significant decrease was observed at the time of fracture,indicating that the microlevel energy release rate was not linearly proportional to the fracture time.These results can help explain the mechanism of cortical bone fractures at the clinical level.

OBJECTIVE To explore wheth er there is a relationship between the judgment results of medical damage liability disputes related to off-label drug use and evidence-based evidence. METHODS By searching for medical damage liability disputes related to off-label drug use up to 2021 on pkulaw.cn ，documents were extracted to record objective factors ，subjective factors and judgment results ；whether there was evidence-based evidence was judged according to Off-label Drug Use List and Evidence-based Evaluation Standards for Off-label Drug Use of Guangdong Pharmaceutical Association ；univariate analysis was adopted to test the relationship between the judgment results and evidence-based evidence. RESULTS A total of 57 cases were included. Cases mainly occurred in the eastern China （63.2％）and tertiary hospitals （64.9％），the main appraisal agency was the appraisal center or institute（61.4％），and the most common type of off-label drug use was overdose drug use （45.6％）. Among the judgment results ， 23 cases（40.4％）of off-label drug use had a causal relationship with medical damage ，most of the responsibility of doctors was secondary responsibility （28.1％），and the actual compensation amount of the most cases were less than 100，000 yuan（54.4％）. There were 25 cases（43.9％）with evidence-based evidence. Univariate analysis found that for off-label drug use the claim amount of the case with evidence-based evidence was significantly higher than that of the case without evidence-based evidence （P＝ 0.040），and there was no significant correlation between evidence-based evidence and the actual compensation amount of the case （P＝0.741），causality determination （P＝0.256），liability type （P＝0.598）or appraisal agency （P≥0.260）. CONCLUSIONS There is no significant correlation between the judgment results of medical damage liability disputes related to off-label drug use and evidence-based evidence ，indicating that there may be certain differences between judicial trials and medical science. The off-label drug use should be regulated by establishing a complete off-label drug use management system and standardizing informed consent procedure for off-label drug use. 1610307322@pku.edu.cn