Objective:To investigate the perinatal prognosis and its impact factors for premature infants with twin-twin transfusion syndrome (TTTS) who were born at ≤34 weeks of gestation.Methods:A retrospective study was conducted on 68 pregnancies of TTTS with gestational age ≤34 weeks at delivery, among them 106 preterm infants (TTTS group) were admitted to the neonatal intensive care unit of the First Affiliated Hospital, Sun Yat-sen University from January 2003 to February 2019. During the same period, another 178 twins without TTTS, congenital malformation, and intrauterine intervention who matched the TTTS group in maternal age (differences within two years) and gestational age (differences within one week) were assigned as non-TTTS group. Perinatal prognosis of TTTS infants born at ≤34 weeks was analyzed by comparing the differences in postnatal early complications and perinatal outcomes (survival time morn than 28 days or not) between the TTTS and non-TTTS groups, recipient and donor twins, mild and severe TTTS infants, and among TTTS infants with different intrauterine interventions. The risk factors for perinatal survival in TTTS infants with gestational age ≤34 weeks were analyzed. Two independent samples t-test, one-way analysis of variance, rank-sum test, Chi-square test, and ordered logistic regression were used for statistical analysis. Results:(1) Among the 68 pregnancies, the overall perinatal survival rate of the neonates was 72.1% (98/136), the double-twin survival rate was 48.5% (33/68), and the rate of at least one survivor was 95.6% (65/68). (2) In the TTTS group, 62 were recipients and 44 were donors. Stage Ⅰ-Ⅱ TTTS was found in 41 cases (mild TTTS group) and stage Ⅲ-Ⅴ in 65 cases (severe TTTS group). (3) The rate of severe brain injury was higher in the severe-TTTS group than those in the mild-TTTS group [9.2% (6/65) vs. 0.0% (0/41), χ 2=4.01, P=0.045]. (4) Gestational age ≤28 weeks ( OR=101.90, 95% CI: 5.07-2 048.37), stage Ⅳ ( OR=14.04, 95% CI: 1.56-126.32) and stage Ⅴ TTTS ( OR=51.09, 95% CI: 3.58-728.81) were independent risk factors for death within 28 days (all P<0.05). (5) Compared with the non-TTTS group, the TTTS group had higher rates of neonatal anemia [51.9% (55/106) vs. 33.1% (59/178), χ 2=9.71], polycythemia [5.7% (6/106) vs. 0.6% (1/178), χ 2=7.18], neonatal persistent pulmonary hypertension [3.8% (4/106) vs. 0.0% (0/178), χ 2=6.81], sepsis [15.1% (16/106) vs. 7.3% (13/178), χ 2=4.40], state Ⅲ or higher retinopathy of prematurity [3.8% (4/106) vs. 0.0% (0/178), χ 2=6.81], congenital cardiac structural abnormality [19.8% (21/106) vs. 0.6% (1/178), χ 2=33.45], heart failure [8.5% (9/106) vs. 0.6% (1/178), χ 2=12.29], and renal insufficiency [14.2% (15/106) vs. 1.1% (2/178), χ 2=20.04] (all P<0.05). Conclusions:Compared with the twin premature infants without TTTS, those with TTTS and ≤34 gestational age were more likely to have cardiac, cerebral, and renal complications. The more severe the TTTS, the higher the incidence of severe brain injury. TTTS preterm infants with gestational age ≤28 weeks and stage Ⅳ or above have high risk of death.

The WRKYs are a group of plant-specific transcription factors that play important roles in defense responses. In this study, we silenced 2 GmWRKY33B homologous genes using a bean pod mosaic virus (BPMV) vector carrying a single fragment from the conserved region of the GmWRKY33B genes. Silencing GmWRKY33B did not result in morphological changes. However, significantly reduced resistances to Pseudomonas syringae pv. glycinea (Psg) and soybean mosaic virus (SMV) were observed in the GmWRKY33B-silenced plants, indicating a positive role of the GmWRKY33B genes in disease resistance. Kinase assay showed that silencing the GmWRKY33B genes significantly reduced the activation of GmMPK6, but not GmMPK3, in response to flg22 treatment. Reverse transcriptase PCR (RT-PCR) analysis of the genes encoding prenyltransferases (PTs), which are the key enzymes in the biosynthesis of glyceollin, showed that the Psg-induced expression of these genes was significantly reduced in the GmWRKY33B-silenced plants compared with the BPMV-0 empty vector plants, which correlated with the presence of the W-boxes in the promoter regions of these genes. Taken together, our results suggest that GmWRKY33Bs are involved in soybean immunity through regulating the activation of the kinase activity of GmMPK6 as well as through regulating the expression of the key genes encoding the biosynthesis of glyceollins.
Glycine max/genetics* ; Disease Resistance/genetics* ; Biological Assay ; Dimethylallyltranstransferase ; Gene Silencing

Objective To explore the clinicopathological and immunohistochemical features of nephrogenic adenoma(NA).Methods Clinical data of NA patients diagnosed in the Department of Pathology,Zhongshan Hospital,Fudan University from July 2016 to October 2022 were collected and analyzed to explore their clinicopathological features.Results A total of 13 NA cases were enrolled.There were 11 males and 2 females.Organs involved:ureter(n=7),bladder(n=5),bladder and ureter(n=1),renal pelvis(n=2).NA patients performed as ureteral stenosis(6/7),rough bladder wall(3/5),and renal pelvis polyp(2/2).The typical microscopical features of NA were tubular(13/13)and papillary(4/13)structures,covered with cuboidal or columnar epithelium(13/13),or a mixed hobnail-spike eosinophilic epithelium(12/13);the interstitium was loose,containing varied amounts of vasculature and inflammatory cells(13/13).Immunohistochemistry revealed specific expressions of CK7,PAX-8,CK19 and CK8.Conclusions NA is a rare neoplasm of the urinary system with unique histological features.NA has the risk of misdiagnosis and over-treatment,and the potential of recurrence and malignant conversion.The diagnosis of NA depends on pathology,and the immunohistochemistry can be helpful for its pathological diagnosis.

Objective To record and analyze the infrared spectra of rubber substrate and film in common pharmaceutical packaging materials,and to provide technical support and reference for the infrared spectra identification of rubber pharmaceutical packaging materials in the national standards system.Methods The infrared spectra of the rubber substrate and film were recorded by the Fourier transform infrared(FT-IR)-attenuated total reflection(ATR)method.The main absorption peaks in the spectra were assigned,the vibration forms were analyzed,and the influencing factors of infrared spectra were discussed.Results The infrared spectra of silicone rubber,polyisoprene rubber,halogenated butyl rubber,ethylene-tetrafluoroethylene copolymer(ETFE),tetrafluoroethylene-hexafluoropropylene copolymer(FEP)and p-xylene were analyzed.Conclusion The production formula of rubber pharmaceutical packaging materials is complex and diverse.Factors such as test methods,rubber product production formula,ATR crystal materials,and silicone oil will affect the results of infrared spectra.

Objective To observe the value of multimodal imaging for diagnosis of cardiac space-occupying lesions.Methods Data of 70 patients with cardiac space-occupying lesions who underwent echocardiography and cardiac CT(CCT)were retrospectively analyzed,among them 35 also underwent cardiac MRI(CMRI).The value of multimodal imaging for diagnosis of cardiac space-occupying lesions were explored according to the results of surgical pathology or clinical diagnosis.Results Among 70 cases,benign tumors were confirmed by surgical pathology in 43 cases,while malignant tumors were confirmed by surgical pathology in 3 cases and clinically diagnosed in 1 case.Meanwhile,non-tumor-occupying lesions were clinically diagnosed in 23 cases,all obviously shrunken after treatments.Among 70 cases,echocardiography correctly diagnosed 57 cases,misdiagnosed 8 cases and unclearly diagnosed 5 cases,with diagnostic accuracy rate of 81.43%(57/70).CCT correctly diagnosed 63 cases,misdiagnosed 4 cases but missed 3 cases,with diagnostic accuracy rate of 90.00%(63/70).CMRI outcomes in all 35 cases were consistent with surgical pathologic results,with diagnostic accuracy rate of 100%(35/35).Conclusion Multimodal imaging might provide objective evidences for diagnosis and treatment of cardiac space-occupying lesions.

The inverse problem of diffuse optical tomography (DOT) is ill-posed. Traditional method cannot achieve high imaging accuracy and the calculation process is time-consuming, which restricts the clinical application of DOT. Therefore, a method based on stacked auto-encoder (SAE) was proposed and used for the DOT inverse problem. Firstly, a traditional SAE method is used to solved the inverse problem. Then, the output structure of SAE neural network is improved to a single output SAE, which reduce the burden on the neural network. Finally, the improved SAE method is used to compare with traditional SAE method and traditional levenberg-marquardt (LM) iterative method. The result shows that the average time to solve the inverse problem of the method proposed in this paper is only 1.67% of the LM method. The mean square error (MSE) value is 46.21% lower than the traditional iterative method, 61.53% lower than the traditional SAE method, and the image correlation coefficient(ICC) value is 4.03% higher than the traditional iterative method, 18.7% higher than the traditional SAE method and has good noise immunity under 3% noise conditions. The research results in this article prove that the improved SAE method has higher image quality and noise resistance than the traditional SAE method, and at the same time has a faster calculation speed than the traditional iterative method, which is conducive to the application of neural networks in DOT inverse problem calculation.
Algorithms ; Neural Networks, Computer ; Tomography, Optical

【Objective】 To study the frequency, Rh phenotypes and molecular & biological background of D-elute (D^el) phenotype in RhD-negative blood donors in Dalian. 【Methods】 A total of 355 serologically RhD-negative samples between November, 2018 and October, 2019 in Dalian Blood Center were collected, and tested for RhC, c, E, e phenotypes using monoclonal antibodies and anti-D adsorption/elution test. DNA was extracted by magnetic bead selection. RHD 1227G>A mutation was detected by melting curve analysis. All RHD exons were sequenced by Sanger sequencing. 【Results】 Among 355 serologically RhD-negative blood donors, 55 (15.5%) were identified as D^el and the remaining 300 cases (84.5%) were true RhD negative. Ccee (45/55, 81.8%) was the predominant Rh phenotype among 55 D^el cases while ccee (210/300, 70.0%) was the most prevalent Rh phenotypes in 300 true RhD negative cases. In 55 D^el cases, 51 (92.7%) had RHD 1227G>A mutation, and the other 4 cases(7.3%) had mutations in other sites. 【Conclusion】 The frequency of D^el was 15.5% in serologically RhD-negative blood donors in Dalian, with Ccee being the most prevalent Rh phenotype and RHD 1227G>A the most common gene mutation.

OBJECTIVETo study the distribution of MICA alleles among ethnic Han Chinese blood donors from Shenzhen and their linkage disequilibrium with HLA-B gene.

METHODSFor 143 randomly selected blood donors, the MICA and HLA-B alleles were determined with a PCR-sequence based typing (SBT) method. Allelic frequency, haplotypic diversity and linkage disequilibrium were analyzed with a Pypop software.

RESULTSThirteen MICA and 35 HLA-B alleles were identified among the 143 blood donors, among which MICA*008:01 had the highest frequency (76/286), whilst MICA*008:01-HLA-B*40:01 and MICA*010-HLA-B*46:01 were the most common haplotypes. No novel allele was identified.

CONCLUSIONThe allele frequencies, haplotype diversities and linkage disequilibrium parameters under a high resolution can facilitate further studies and applications of the MICA and HLA-B genes.

OBJECTIVETo detect potential mutation of EXT1 gene in a pedigree affected with multiple osteochondroma and explore its pathogenic mechanism.

METHODSThe coding regions and their flanking sequences of the EXT1/EXT2 genes were subjected to PCR amplification and Sanger sequencing. Suspected mutations were verified by excluding possible single nucleotide polymorphisms and bioinformatics analysis. Transcripts of the EXT1 gene in the proband were analyzed by TA clone-sequencing, with its abundance compared with that of healthy controls.

RESULTSDNA sequencing has identified in the proband a novel heterozygous point mutation (c.1164+1G to A) at the 5'splice sites of intron 3 of the EXT1 gene. The same mutation was not found in the healthy controls. Bioinformatics analysis indicated that the mutation is highly conserved and can lead to skipping of exon 3 or aberrant splicing. TA clone-sequencing indicated that the numbers of transcripts with skipping of exon 3 has significantly increased in the proband (< 0.05) compared with the controls.

CONCLUSIONThe c.1164+1G to A mutation has resulted in skipping of exon 3 in a proportion of EXT1 gene transcripts. As the result, the number of transcripts with tumor suppressing function is relatively reduced and has ultimately led to the tumors.

Adult ; Base Sequence ; Child ; Exostoses, Multiple Hereditary ; genetics ; Female ; Humans ; Male ; Molecular Sequence Data ; N-Acetylglucosaminyltransferases ; genetics ; Point Mutation ; RNA Splice Sites ; RNA Splicing

BACKGROUND: Due to the polymorphism of HLA, a large number of ambiguities have been generated by conventional HLA typing techniques, and confirmed stereotypes of ambiguous results based on group-specific haploid full-length typing are rarely reported.OBJECTIVE: To analyze the accuracy of HLA-typing ambigulity based on group-specific haploid full-length sequencing. METHODS: The low-resolution results were used as the starting point for two ambiguous samples. Sanger sequencing (PCR-SBT) based on haploid full-length was performed after group-specific amplification. RESULTS AND CONCLUSION: One case showed a new A*02:03:01 allele, which was found a mutation in NT817 from C to T in comparison with A*11:01:01:01. The other case indicated another new C*07:02:01:01, which was found a mutation in NT879 from A to G in comparison with C*08:01:01. In conclusion, these results indicate that the group-specific haploid full-length sequencing method can be used to accurately classify HLA alleles and to discover new alleles.