[Objective] To explore the accuracy and feasibility of non-invasive prenatal diagnosis of fetal RhE genotype using cell-free fetal DNA (cff-DNA) from maternal peripheral blood. [Methods] A total of 134 pregnant women with single fetuses and RhE-negative blood group were selected from our hospital from November 2023 to August 2024. Free DNA extraction kit was used to extract free DNA from peripheral blood of pregnant women, and the RhE blood group genotype of free DNA was detected by real-time fluorescent quantitative PCR (RT-qPCR). If the qPCR amplification signal of the sample was negative, the methylated RASSF1A gene was amplified, and the positive amplification result was used as a sign of successful extraction of cff-DNA. Serological microcolumn gel method was used to detect the phenotype of RhE blood group in neonatal peripheral blood. [Results] Among the 134 maternal peripheral blood samples, the cff-DNA detection of RhE blood group phenotypes was consistent with the RhE blood group genotyping of neonatal peripheral blood in 133 cases, including 90 cases of Rhee genotype and 43 cases of RhE genotype, with diagnostic concordance rate of 99.3%, sensitivity of 97.7%, specificity of 100%, youden index of 0.977, area under ROC curve of 0.995, the Kappa value of 0.983, positive predictive value of 100%, and negative predictive value of 98.9%. The sample of 1 case failed to be detected. After the amplification of methylated RASSFIA gene, it was confirmed that the reason for the failure was that no cff-DNA was extracted from the sample. The diagnostic concordance rates of the first, second and third trimesters were 93.8% (15/16), 100% (51/51) and 100% (67/67), respectively. Fisher's exact test method was used to calculate the P value, which was P>0.05, indicating that there was no statistical significance in the difference of diagnostic concordance rate among the three pregnancy periods, and there was no difference in the detection concordance rate of this method in different pregnancy periods. [Conclusion] The use of cff-DNA in maternal peripheral blood for the detection of fetal RhE blood group genotype is an accurate and highly feasible non-invasive prenatal diagnostic method, which is helpful for the clinical diagnosis of fetal and neonatal hemolytic disease caused by anti-E antibody.

Objective To investigate the expression levels of serum microRNA-33a-5p(miR-33a-5p)and Sirtuin6(SIRT6)in patients with esophageal cancer,and their relationship with clinical pathological characteristics and prognosis.Methods From March 2018 to March 2020,123 newly diagnosed patients with esophageal cancer(esophageal cancer group)and 94 patients with esophagitis(benign lesion group)admitted to our hospital were collected as the research subjects,another 72 healthy individuals who underwent physical examination were regarded as the health control group;the clinical and pathological characteristics of esophageal cancer patients were collected.Real-time fluorescence quantitative PCR method was applied to detect the expression of miR-33a-5p and SIRT6 mRNA in three groups of serum,the relationship between the expression levels of serum miR-33a-5p and SIRT6 mRNA in esophageal cancer patients and clinical pathological characteristics was analyzed;TargetScanHuman website was applied to predict the targeting relationship between miR-33a-5p and SIRT6;Pearson method was applied to analyze the correlation between serum miR-33a-5p and SIRT6 mRNA expression levels in esophageal cancer patients;Kaplan-Meier method was applied to analyze the relationship between the expression levels of serum miR-33a-5p and SIRT6 mRNA in esophageal cancer patients and their prognosis;Cox regression was applied to analyze the prognostic factors of esophageal cancer patients.Results The expression level of serum miR-33a-5p in the esophageal cancer group was greatly higher than that in the benign lesion group and healthy control group(P＜0.05),the expression level of serum SIRT6 mRNA was greatly lower than that of the benign lesion group and healthy control group(P＜0.05);TargetScanHuman website predicts that there was a targeted binding site between miR-33a-5p and SIRT6,and there was a negative correlation between serum miR-33a-5p and SIRT6 mRNA expression levels in esophageal cancer patients(r=-0.468,P＜0.05);the expression levels of serum miR-33a-5p and SIRT6 mRNA in esophageal cancer patients were related to TNM staging,lymph node metastasis,and degree of differentiation;the 3-year survival rate of esophageal cancer patients with low expression of serum miR-33a-5p and high expression of SIRT6 mRNA was greatly higher than that of esophageal cancer patients with high expression of miR-33a-5p and low expression of SIRT6 mRNA(P＜0.05);Cox regression analysis showed that high expression of miR-33a-5p and low expression of SIRT6 mRNA were independent risk factors for death in esophageal cancer patients(P＜0.05).Conclusion The expression level of serum miR-33a-5p in esophageal cancer patients is greatly increased,while the expression level of SIRT6 mRNA is greatly reduced;the expression levels of both are closely related to the clinical pathological characteristics and prognosis of esophageal cancer patients.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To investigate the feasibility of deep learning radiomics model in the prediction of neoadjuvant chemotherapy (NAC) response in breast cancer based on ultrasound images at an early stage.Methods:Between January 2018 and June 2021, 218 patients with breast cancer who underwent NAC were enrolled in the retrospective study. All patients received a full cycle of NAC before surgery and underwent standard ultrasound examination before NAC and after the second cycles of NAC. Of all the patients, 166 patients came from institution 1 (the First Affiliated Hospital of Nanjing Medical University) were allocated into a primary cohort.Based on the architecture of Resnet 50 convolutional neural, a deep learning prediction model was built.Further validation was performed in an external testing cohort ( n=52) from institution 2 (General Hospital of Eastern Theater Command, PLA). The clinical model was constructed using independent clinical variables. To evaluate the predictive performance, areas under the curve (AUCs) of these models and two radiologists were compared by using the DeLong method. Results:The Resnet 50 model predicted the response of NAC with accuracy. The deep learning model, achieving an AUC of 0.923 (95% CI=0.884-0.962) in the primary cohort and an AUC of 0.896 (95% CI=0.807-0.980) in the test cohort, outperformed the clinical model and also performed better than two radiologists′ prediction (all P<0.05). Furthermore, the two radiologists achieved a better predictive efficacy (AUC 0.832 and 0.808 for radiologists 1 and 2, respectively) when assisted by the DL model (all P<0.01). Conclusions:The deep learning radiomics model is able to predict therapy response in the early-stage of NAC for breast cancer patients, which could guide clinicians and provide benefit for timely treatment strategy adjustment.

ObjectiveTo explore the influence of the core symptoms of attention deficit hyperactivity disorder （ADHD） on behavioral problems of children with ADHD propensity， so as to provide references for early identification and targeted intervention for children with ADHD propensity. MethodsFrom July to August 2021， 25 children with ADHD propensity were screened as the ADHD propensity group， and 25 children matched for age， gender and grade were included as the normal group in an elementary school in Guangzhou. ADHD core symptoms were assessed by the Chinese version of the Swanson Nolan and Pelham， version IV-parent form for ADHD （SNAP-IV）， and behavioral problems were assessed by Questionnaire-Children with Difficulties （QCD） and Conners Parental Symptom Questionnaire （PSQ）. Spearman correlation analysis was used to examine the correlation between ADHD core symptoms and QCD and PSQ scores， and hierarchical linear regression analysis was used to explore the effect of ADHD core symptoms on behavioral problems. Results① The differences between the groups showed that both attention deficit and hyperactivity-impulsivity factor scores were higher in the ADHD propensity group than those in the normal group （t=7.771， 6.726， P<0.01）. ② Correlation analysis showed that the attention deficit factor score was negatively correlated with QCD total score （r=-0.440， P<0.05）， and positively correlated with the learning problem factor score of PSQ （r=0.457， P<0.05）. The score of hyperactivity-impulsivity was negatively correlated with score of anxiety factor in PSQ （r=-0.457， P<0.05）， and positively correlated with impulse-hyperactivity factor score （r=0.552， P<0.01）. ③ Hierarchical linear regression analysis showed that the attention deficit factor score negatively predicted the total score of QCD （B=-0.682， P<0.05， R²=0.468）. The hyperactivity-impulsivity factor score had a negative predictive effect on the anxiety factor score of PSQ （B=-0.048， P<0.05， R²=0.367）， and had a positive predictive effect on the impulsivity-hyperactivity factor score （B=0.077， P<0.01， R²=0.424）. ConclusionChildren with ADHD propensity have significant attention deficit symptoms， hyperactivity-impulsivity symptoms and behavioral problems， and the attention deficit may be the main cause of their daily behavioral problems， while hyperactivity-impulsivity may be the main cause of their impulsive-hyperactivity problems.

OBJECTIVE：To study the skin irritation and se nsitization of domestic generic drug Clobetasone butyrate cream ， and to compare it with commercial drug （original drugs ）. METHODS ：The skin irritation test was conducted on rabbits. Totally 24 rabbits were randomly divided into test preparation intact skin group ，test preparation abraded skin group ，commercial drug intact skin group and commercial drug abraded skin group ，with 6 rabbits in each group. 0.5 mL test preparation or commercial drug was administered to the left side of intact or abraded skin and the same amount of excipient on the right side of each rabbit twice a day for consecutive 7 days. The irritation of the drug to the rabbit skin was observed ，and the erythema and edema of the skin were scored；the skin of administration site was taken at 72 h after last administration and the end of 7 d after drug withdrawal for histopathological examination. The skin sensitization test （Buehler test ）was carried out on guinea pigs. Totally 60 guinea pigs were randomly divided into test preparation group （n＝20），commercial drug group （n＝20），positive control group （n＝10）and excipient control group （n＝10）. 0.2 mL test preparation or commercial drug was administrated to the left side of the rib abdomen skin of each guinea pig at the 0，7th，14th day to induce model ，and an equal amount of corresponding preparation was administered to the right side in the same way at the 28th day for stimulation. Hypersensitive response such as erythema and edema were observed and scored at 24 h and 48 h after the stimulation. The incidence of hypersensitive response was then calculated. RESULTS：In skin irritation test of rabbits ，no erythema and edema was caused by the test preparation or commercial drug on intact skin of rabbits ；scores of skin irritation was 0；there was no dermal irritation. Both test preparation and commercial drug caused transient slight erythema on abraded skin of a few rabbits ；scores of intact and abraded skin irritation were 0-0.33；there was no dermal irritation. There was no statistical significance among groups. No dermal pathological changes were observed. In skin sensitization test of guinea pig ，no hypersensitive response such as erythema and edema was found on the skin of guinea pigs in both test preparation and commercial drug groups ；both score and the incidence of hypersensitive response were 0. Compared with excipient control group ，there was no statistical significance of average score and the incidence of hypersensitive response in test preparation group and commercial drug group. CONCLU- SIONS：In skin irritation test of rabbits and skin sensitization test of guinea pigs ， the evaluation results of generic Clobetasone butyrate cream are the same as those of the original drug. It has no irritation to the skin of rabbit ，and no sensitization to the skin of guinea pigs.

Objective To investigate the expression of c-Myc in gastric mucosa associated lymphoid tissue (MALT) lymphoma and its implications on prognosis.Methods The clinical data of 79 patients hospitalized in our hospital from 2009 to 2015 were retrospectively analyzed.In these patients,38 patients were low-grade MALT lymphoma,20 patients were high-grade MALT lymphoma,21 patients were diffused large B cell lymphoma (DLBCL).Real-time PCR was used to detect the levels of c-Myc expression in gastric MALT tissues and pair-matched adjacent normal tissues.The relationship between the expression of c-Myc and prognosis of patients was evaluated combing with the clinical data.Results Compared with the normal tissues,the expression levels of c-Myc protein were 15.7％ (6/38),25％ (5/20) and 28.5％ (6/21)in patients with low-grade MALT lymphoma,high-grade MALT lymphoma,and DLBCL.The relative expression levels of cMyc mRNA were gradually elevated in low-grade MALT lymphoma,high-grade MALT lymphoma and DLBCL.The tumor size and depth of invasion can influence the expression level of c-Myc.Survival analysis found that the overall survival rates and relapse-free survival rates were lower in patients with c-Myc positive expression than those of patients with negative expression (P ＜ 0.05).Conclusion C-Myc plays a key role in the malignant transformation of gastric MALT lymphoma.

Objective To evaluate the application of atherogenic index of plasma (AIP) and intima-media thickness of carotid artery (CA-IMT) in renal arteriolar sclerosis patients with chronic renal failure.Methods One hundred and twenty nine patients with chronic renal failure patients underwent ultrasound-guided percutaneous renal biopsy from October 2013 to June 2014,the biopsy results showed that renal arteriolar sclerosis was identified in 72 patients (atherosclerosis group) and no renal arterioles sclerosis was detected in 57 patients (non-atherosclerosis group);71 healthy adults were enrolled in the study as controls.The age,height,body weight,systolic and diastolic blood pressure,the indexes of blood lipid and renal function were documented and compared among three groups.The correlation of AIP and CBMmax of common carotid artery and carotid bifurcation with blood lipid level and renal function was analyzed.Results There was significant difference in body weight among patients with atherosclerosis [(70.77 ± 14.27) kg],without atherosclerosis [(60.63 ± 12.12) kg] and the controls [(64.20 ± 8.13) kg] (t =3.071,3.391,all P ＜ 0.05).The TG [(2.43 ± 1.61) mmol/L vs.(1.02 ± 0.37) mmol/L],TC [(7.40 ± 8.80) mmol/L vs.(4.53 ±0.67)mmol/L],LDL-C[(4.40 ±2.13) mmol/L vs.(2.85 ±0.70) mmol/L],AlP[(0.15 ± 0.351) vs.(-0.127 ± 0.184)] of the atherosclerosis group were higher than those of control group (t =5.975,2.252,2.614,-5.467,all P ＜ 0.05).The HDL-C of atherosclerosis group was lower than that of control group [(0.78 ±0.16) mmol/L vs.(1.29 ±0.21) mmol/L,t =4.750,P ＜0.05].The Scr[(117.24 ± 94.27) mmol/L vs.(64.16 ± 13.42) mmol/L],BUN [(6.73 ± 3.58) mmol/L vs.(4.66 ± 1.08) mmol/L] of the atherosclerosis group were higher,and the GFR was lower [(65.60 ±23.00)ml · min-1 · 1.73 m-2 vs.(124.78 ± 24.35)ml · min-1l.73 m-2,t =5.118] than those of control group (t =4.730,4.702).The Scr of the atherosclerosis group was higher,and the GFR was lower [(65.60 ± 23.00) ml · min-1 · 1.73 m-2 vs.(95.60±53.00)ml · min-1 · 1.73 m-2,t =3.514] than those of the non-atherosclerosis group [(117.24 ± 94.27) mmol/L vs.(71.35 ± 42.18) mmol/L,t =3.690].There were positive correlation between TG and LDL-C (r =0.828,0.323,P ＜ 0.05) and negative correlation between AIP and HDL-C (r =-0.489,P ＜0.05).There was positive correlation of CBMmax with Scr,BUN and AIP (r =0.394,0.289,0.528,all P ＜ 0.05),and negative correlation between CBMmax and GFR (r =-0.277,P ＜ 0.05).Conclusion Body weight,GFR,AIP and CBMmax are useful indicators in evaluation of renal arteriolar sclerosis in patients with chronic renal failure.AIP is a sensitive index for abnormal blood lipid level.AIP and CBMmax are important risk factors in chronic renal failure patients with renal arteriolar sclerosis.

BACKGROUND:Urine-derived stem cels are most likely to come from the kidney tissue, and therefore, these cels are more adaptable to kidney microenvironment, providing a new option for the treatment of kidney diseases.
OBJECTIVE: To explore the therapeutic efficacy of human urine-derived stem cels on chronic nephropathy rats.
METHODS:The fresh urine samples of healthy people were colected, and then human urine-derived stem cels were extracted and cultured in vitro. Twenty Sprague-Dawley rats were used to prepare chronic nephropathy models, and given injection of human urine-derived stem cel suspension (experimental) or normal saline (control) into the renal cortex, respectively. Another 10 healthy rats were used as controls. Therapeutic effects on renal function were assessed by detection of serum creatinine level and glomerular filtration rate in the three groups. The kidney tissues of rats were taken and observed histomorphologicaly in each group.
RESULTS AND CONCLUSION: Human urine-derived stem cels were found to remarkable improve rat’s renal function as wel as reduce the histomorphological changes in the kidney tissues of rats. Compared with the control group, the serum creatinine level was decreased while the glomerular filtration rate was increased significantly in the experimental group; CD68 expression and infiltration of interstitial inflammatory cels were also markedly reduced in the experimental group. To conclude, human urine-derived stem cels can improve the renal function of chronic nephropathy rats.

Objective To evaluate the influence of SOCS3 conditional ablation in macrophage polarization and functional recovery after spinal cord injury in mice.Methods The Socs3fVfl Nes cre mice were used to obtain nervous-system-restricted SOCS3 deletion models as experimental group (n=40),and littermate Socs3fl as control group (n=40).Mice (n=35) in each group were underwent T10 spinal cord compressing injury (the left 5 were as sham-operated controls);on the third,7th and 14th d of injury,injured tissues from 15 mice of each group were harvested;immunofluorescence was performed to detect the changes of M2a and M2c phenotypic marker Arginase1 and M1 and M2b phenotypic marker CD86;on the first,third,7th and 14th d of injury,injured tissues from the left 20 mice from each group were harvested;fluorescent quantitative real time-PCR was used to detect the macrophage phenotypic markers (Arginasel,CD206,iNOS and CD32),SOCS3 and STAT3 gene expressions;the above data from the sham-operated controls were used as base data.BMS scale was used to evaluate the motor functions of posterior limbs and bodies on the first,third,7th and 14th d of injury.Results The behavioral assessments of the experimental group were superior on the third,7th and 14th d of injury as compared with those of control group (P＜0.05).Immunofluorescence showed that as compared with the controls,the Socs3fVfl Nes cre mice had a predominance in distribution and population of Arginasel-positive macrophages in spinal cords on the third,7th and 14th d of injury;in contrast,the control group showed superiority of CD86 expression on the 7th and 14th d,with significant differences (P＜0.05).Similarly,fluorescent quantitative real time-PCR analysis demonstrated that gene expression of A rginase I on the 7th d of injury,that of CD206 on the 7th and 14th d of injury,and that of STAT3 on the third,7th,and 14th d of injury were significantly increased,whereas that of CD32 on the 14th d of injury and SOCS3 on the first and third d of injury were significantly reduced in the experimental group,as compared with those in the control group (P＜0.05).Conclusion SOCS3 conditional knockout can promote the formation of M2 macrophage,as well as good functional recovery after spinal cord injury in mice.