[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 explore the clinical effect of PRP on refractory ulcer of diabetes foot on the basis of routine treatment. 【Methods】 Sixty-four patients who suffered from diabetes foot and treated in our hospital from January to December 2020 were divided into the routine treatment group (44 cases) vs PRP plus routine treatment group (20 cases, using liquid or gel PRP for diversified treatment) according to a simple random sampling method. The general conditions of the two groups were evaluated to compare the wound surface, wound healing rate, treatment time, wound healing speed rate, adverse reactions and healing conditions after the treatment. 【Results】 The wound surface[0.05(0.00, 0.70)vs 0.35(0.00, 4.54)], wound healing rate[0.99(0.84, 1.00)vs 0.80(0.26, 1.00)] and wound healing speed rate[0.16(0.04, 0.27)vs 0.06(0.01, 0.18)] in PRP group were significantly higher than those in routine treatment group (P<0.05). The treatment duration[18.00(8.75, 24.75)vs 17.00(9.25, 28.00)] between the two groups was not statistically significant (P>0.05), so was the adverse reactions to treatments[0(0/20)vs 2.27(1/44)](P>0.05). The response rate[100(20/20)vs 61.36(27/44)] of PRP group was significantly better than that of routine group, and the difference was statistically significant (P<0.05). 【Conclusion】 The therapeutic effect of PRP group was significantly superior to that of routine treatment group.

The successful retrieval of ancient mitochondrial DNA (mtDNA) from Neanderthals provides powerful experimental evidence that clarifies the arguments between the out-of-Africa and multiregional models of evolution. However, the lack of nuclear DNA from Neanderthal fossils and mtDNA of early modern human fossils dating back to approximately the same time in the Pleistocene constitutes a limitation that may compromise the significance of mtDNA phylogenetic analysis. In this report, we introduce a mitochromic analysis using Neanderthal mtDNA as a foreign transgene and humans as a naturally occurring transgenic species. Forty Neanderthal mtDNA retrievable nuclear fragments were identified by blasting human genome data with Neanderthal mtDNA. Five of the 40 fragments exhibited higher correlation with Neanderthal mtDNA than those with modern human mtDNA. Furthermore, these five nuclear fragments harbor Neanderthal mtDNA-unique haplotypes. Based on the 98%+ identity between Neanderthal and modern human mtDNA when compared by groups, we suggest that some of the modern human nuclear fragments retrieved using Neanderthal mtDNA may aid in decoding Neanderthal genetic information, and also may simultaneously demonstrate a close genetic evolutionary relationship between modern humans and Neanderthals.

ObjectiveTo investigate the potential mechanism of serum N-glycan alterations in patients with hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) by measuring serum N-glycan profile and comparing glycosyltransferase gene expression between HCC tissue and adjacent tissue. MethodsThe samples of HCC tissue, adjacent tissue, and normal liver tissue were collected from 34 patients with HBV-related HCC who were admitted to Chinese PLA General Hospital, and serum samples were also collected. Among these 34 patients, 8 were randomly selected and their serum samples were established as HCC experimental group, and the serum samples of 20 healthy adults were established as control group. DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis was used to analyze serum N-glycan profile in the HCC experimental group and the control group. Quantitative real-time PCR was used to measure the mRNA expression of 8 glycosyltransferase genes (FUT3, FUT4, FUT6, FUT7, FUT8, Gn-TIII, Gn-TIVa, and Gn-TV) in the HCC tissue and adjacent tissue of 34 patients with HBV-related HCC, and Western blot was used to measure the expression of corresponding proteins. The independent samples t-test was used for comparison of continuous data between two groups. ResultsCompared with the control group, the HCC experimental group had a significant increase in the abundance of N-glycan peak9 (NA3Fb) in serum(t=-2.514,P＜0.05). There were significant differences in the mRNA expression of FUT8, Gn-TIVa, and Gn-TV between HCC tissue and adjacent tissue, and the mRNA and protein expression levels of FUT8 and Gn-TV in HCC tissue were significantly higher than those in adjacent tissue (FUT8 mRNA: 1.50±0.34 vs 0.65±0.11, t=-2.354，P=0.022; Gn-TV mRNA: 3.57±0.64 vs 1.33±016, t=-3.384,P=0001; FUT8 protein: 0.70±0.11 vs 0.083±0.017, t=9.555,P=0.001; Gn-TV protein: 1.33±0.19 vs 0.60±0.15, t=5.097,P=0.007). The mRNA expression level of Gn-TIVa in HCC tissue was significantly higher than that in adjacent tissue (2.90±0.47 vs 1.68±0.19, t=-2.403,P=0.019), but there was no significant difference in the protein expression level of Gn-TIVa between HCC tissue and adjacent tissue (052±0.24 vs 0.24±0.11,t=1.833, P=0.141). The changes of glycosyltransferase gene expression in HCC tissue were consistent with the alteration of serum N-glycan profile. ConclusionSerum N-glycan alterations in patients with HBV-related HCC may be closely associated with the upregulated expression of the glycosyltransferase genes FUT8, Gn-TIVa, and Gn-TV in HCC tissue.

Objective To evaluate the alveolar bone thickness and root length changes of anterior teeth with cone-beam computed tomography(CBCT).Methods CBCT scans were taken for 12 skeletal Class Ⅲ patients who accepted the improved corticotomy(IC) procedures during pre-surgical orthodontics.The CBCT data in T1(the maxillary dental arch was aligned and leveled) and T2(extraction space closure) were superimposed and the alveolar bone thickness at root apex level and root length measurements were done.Results From T1 to T2,the buccal alveolar bone thickness for the upper lateral incisors increased from (1.89±0.83) to (2.47± 1.02) mm (P＜0.05),and for central incisors and for canines from (2.32±0.71) to (2.68 ± 1.48) mm and from (2.28 ± 1.08) to (2.41 ± 1.40) mm,respectively.According to Sharpe Grading System,the root resorption grade for 69 teeth of 72 was located in Grade 1,two teeth in Grade 2,one tooth in Grade 3.Conclusions The improved corticotomy had the potential to increase the buccal alveolar bone thickness and the root resorption in most teeth was in Grade 1 according to Sharpe grading system.

A 77-year-old man was admitted to our hospital at July 5th,2010 with an unexplained massive pericardial effusion for 10 years.With dyspnea for one month and normal vital signs without pulsus paradoxus,other physical examination included a small amount of moist rale,normal heart sounds,jugular vein engorgement,positive hepatojugular reflux,hepatosplenomegaly and pitting edema of the extremities.The patient had a complex past history with lymph node tuberculosis,primary artertial hypertension,polycythernia vera,chronic renal insufficiency and hypothyroidism (Hashimoto's thyroiditis),and moreover,received a high dose radiation of 31p in 1967. Family history is negative.The patient had no cardiac tamponade or pericardial constriction during 10 years,he received pericardiocentesis twice,and pericardial effusion was exudative with a high proportion of monocyte.There was no evidences of tuberculosis infection,hypothyroidism,malignant tumor,severe heart failure,uremia,trauma,severe bacterial or fungus infection,chronic myeloid leukemia or bone marrow fibrosis during the admission. The patient refused anti tuberculosis,indwelling catheter drainage or surgical therapy.In this rare case,the aetiology of chronic massive pericardial effusion is most probably chronic idiopathic recurrent pericarditis.

Objective:To investigate the relationship between fester capacity and clinical efficacy on allergic asthma treated by scarring moxibustion.Methods:The patients were randomly divided into scarring moxibustion 9-cone group,3-cone group and 6-cone group.The biggest area,scab-lost time,first-festering time and scarring time of moxibustion sore,and the clinical efficacy in each group was observed.Results:The biggest post-moxibustion sore area in 9-cone group was larger than that in 3-cone group,but had no difference in comparison with 6-cone group;the scab-lost time,first-festering time and scarring time of post-moxibustion sore in each group were of no difference.The clinical efficacy was better in 9-cone group than in 3-cone group,but had no difference in comparison with the 6-cone group.Conclusion:Fester capacity in treating allergic asthma by scarring moxibustion is related to the clinical efficacy and definite fester capacity is the key to good results on allergic asthma.

cholesterol 7α-hydroxylase gene ( CYP7A 1 ) plays a key role in the catabolism of cholesterol into bile acids. To investigate whether the A-204C polymorphism in CYP7A1 gene affects the gene expression,using luciferase as the reporter gene, four recombinants were constructed by inserting forward or reverse sequence with A or C allele at the polymorphism site into the promoter-less vector pGL3-basic. The constructs were then transfected into four cell lines and the luciferase activity of each expression vector was examined by dual luciferase reporter gene assay system. The results showed that activities of the forward sequence of both genotypes were higher than that of reverse sequence. Promoter activity of the recombinants with A allele was about one third lower than that with C allele. According to the analysis with TRANSFAC database, there may exist a Zic3 binding site when there is the C allele at -204. Our study indicates that the A-204 C polymorphism in CYP7A1 promoter region decreases its promoter activity and thus represses the gene expression, possibly due to the lack of a potential Zic3 binding site.

OBJECTIVETo analyze the pattern and prevalence of partial copy deletion of deleted-in-azoospermia (DAZ) gene in the azoospermia factor C(AZFc) region of patients with idiopathic azoospermia or severe oligozoospermia.

METHODSsY581 and sY587 in DAZ gene region were analyzed by polymerase chain reaction-restriction length polymorphism(PCR-RFLP) for its deletion in 197 patients with azoospermia, 166 patients with severe oligozoospermia, and 210 fertile men as controls.

RESULTSDeletion of both DAZ1 and DAZ2 was detected in 18 patients with azoospermia and 10 with severe oligozoospermia, and the prevalence was 9.1% and 6.0% respectively. There was significant difference in deletion rate between the cases and controls.

CONCLUSIONThe frequency of partial copy deletion of DAZ gene in Chinese idiopathic azoospermia or severe oligozoospermia patients is much higher than that of fertile controls, suggesting that the deletion of DAZ1/DAZ2 may be one of the important genetic etiological factors of spermatogenesis damage. The pattern and prevalence of DAZ partial copy deletion are similar to those of Caucasians populations, and detection of DAZ gene partial copy deletion by PCR-RFLP may be adopted as an additional clinical gene diagnostic measure after AZF microdeletion detection.

Azoospermia ; complications ; genetics ; Chromosomes, Human, Y ; genetics ; Deleted in Azoospermia 1 Protein ; Gene Deletion ; Humans ; Infertility, Male ; etiology ; genetics ; Male ; Models, Genetic ; Polymerase Chain Reaction ; RNA-Binding Proteins ; genetics

By analyzing the traditional educational mode and subject of the higher medical education, the basic method and emphasis of the experimental class of the clinical medicine educational innovation have been established, and the general frame of the educational innovation content has been constructed, the executive essential point and point in the content of the educational innovation have been made. After several years of the practice and positive exploration, the class has achieved success.