[Construction of microRNA-21 terahertz metamaterial sensing method based on duplex-specific nuclease triggered rolling circle amplification].

Objective: To construct of a terahertz metamaterial sensing method for detection of microRNA-21 (miRNA-21) based on duplex-specific nuclease triggered rolling circle amplification. Methods: First, a THz metamaterial sensing method was constructed, and verified by polyacrylamide gel electrophoresis and zeta potential; after optimizing the detection conditions of the sensor, different concentrations of miRNA-21 and other different miRNAs were detected. And this biosensor was compared with other miRNA detection methods; finally, the recovery rate of the biosensor was evaluated. Results: Under the optimal experimental conditions, through the dual signal amplification strategy of duplex-specific nuclease (DSN) cycle recognition and rolling circle amplification (RCA), the THz metamaterial sensor has a response range of 10 fmol/L to 10 nmol/L to the target miRNA-21, with a detection limit of 8.49 fmol/L. And the biosensor has good specificity with the ability to recognize the target miRNA-21 from a variety of microRNAs. And the experiment with recovery rate from 94.33% to 115.33% has been further verified in commercial human serum samples. Conclusion: The terahertz biosensor can achieve highly sensitive and specific detection of the target miRNA-21, which proves the potential for label-free diagnosis and early warning of miRNA-related diseases.

[A novel compound heterozygous mutation in MYSM1 gene in a 1-month-old girl: a bone marrow failure syndrome 4 family survey and literature review].

Objective: To report the clinical manifestations and total exon detection results of one case of MYSM1 gene complex heterozygosity mutation of bone marrow failure syndrome 4 and the results of total exon detection of her family to provide a case phenotype for the early diagnosis of bone marrow failure syndrome 4. Methods: A 1-month-old girl with severe anemia was sequenced with trio-WES. Similarly, the family was also sequenced with tribe-WES to confirm the molecular diagnosis. BWA, GATK, and other software were used for annotation analysis of sequencing results. After polymerase chain reaction, Sanger sequencing was performed by ABI3730 sequencer to verify the target sequence. Moreover, the verification results were obtained by the sequence analysis software. The clinical diagnosis of this girl was reported and the relevant pieces of literature were reviewed. Results: The girl presented with pancytopenia, polydactylism, nonspecific white matter changes, and cysts. However, CD3(-)CD19(+) B decreased. The child was identified with MYSM1 complex heterozygous mutation by whole-exome sequencing, NM_001085487.2:c.1607_c.1611delAAGAG and c.1432C>T, which was respectively inherited from his parents. Genealogy verification confirmed that the c.1432C>T mutation carried by the father was from the grandfather (father's father) , whereas the c.1607_c.1611delAAGAG mutation carried by the mother was from the grandfather (mother's father) , whereas the grandmothers, aunts, and uncle did not carry the mutation. The child was diagnosed with BMFS4 combined with clinical phenotypic and molecular genetic findings. Conclusion: This case provides a case phenotype for the early diagnosis of BMFS4 and extends the pathogenicity variation and phenotype spectrum of the MYSM1 gene. The newly discovered pathogenic variant of MYSM1 c. 1607_c.1611delAAGAG has not been reported at home or abroad.

[A comparative study of lacrimal magnetic resonance hydrography and lacrimal endoscopy examination in the diagnosis and treatment of lacrimal duct obstructive diseases].

OBJECTIVE: To evaluate the diagnostic value and treatment guidance of lacrimal magnetic resonance hydrography (LMRH) and lacrimal endoscopy examination in lacrimal duct obstruction. METHODS: A retrospective analysis of clinical and imaging data of 59 patients with epiphora who had LMRH examination in Tongji Hospital during June 2013 and January 2014. Multiplanar reconstruction (MPR) and maximum intensity projection (MIP) were used to process the three dimensions T2-weighted images (T2WI). The size of lacrimal sac, lacrimal mucosal lesions and the obstructed plane of nasolacrimal duct were observed. The lacrimal irrigation results were used as gold standard. The sensitivity, specificity, accuracy of LMRH in diagnosis of lacrimal duct obstructive diseases and the consistency between the two methods were analyzed. In addition, 22 cases had lacrimal endoscopy examination in less than half month after MRD. The results of lacrimal endoscopy were compared with LMRH images. The treatment method was made according to the results of LMRH and lacrimal endoscopy. RESULTS: According to the results of lacrimal irrigation, among 78 eyes of 59 patients, 2 eyes were diagnosed as lacrimal canalicular obstruction (2.6%, 2/78), 8 eyes were diagnosed as nasolacrimal duct stenosis (10.3%, 8/78), 24 eyes were diagnosed as nasolacrimal duct obstruction (30.8%, 24/78), 44 eyes were diagnosed as nasolacrimal duct obstruction accompanied with chronic dacryocystitis (56.4%, 44/78). The other 40 eyes were negative controls. LMRH had a high degree of consistency with lacrimal irrigation in diagnosis of lacrimal duct obstructive diseases. The value of Kappa was 0.963 (P= 0.026). The sensitivity of MRD in diagnosis of lacrimal duct obstructive diseases was 97.4%, the specificity was 100%, the accuracy was 98.3%, the positive predictive value was 100% and the negative predictive value was 95.2% . According to 40 eyes of the control group, the mean value of the maximum cross-sectional area of the lacrimal sac was: (10.9 ± 0.4) mm(2). Twenty-two eyes underwent lacrimal endoscopy examination and the endoscopic findings were consistent with LMRH diagnosis. The lesions in the lacrimal duct displayed more clearly and intuitively than the LMRH, while LMRH had its unique advantages in showing the size of lacrimal sac, the mucosal thickness of lacrimal duct, large foreign bodies and lesions around the lacrimal duct. According to the results of LMRH and lacrimal endoscopy, 2 eyes of canalicular obstruction, 8 eyes of nasolacrimal duct stenosis, 20 eyes of nasolacrimal duct obstruction underwent lacrimal probing and stent implantation. Four eyes of nasolacrimal duct obstruction had drug treatment under lacrimal endoscopy. Thirty-eight eyes of chronic dacryocystitis underwent endonasal dacryocystorhinostomy. The other 6 eyes of chronic dacryocystitis underwent stent removal combined with endonasal dacryocystorhinostomy. CONCLUSIONS: LMRH is a noninvasive and reliable method to examine the lacrimal duct obstruction. It can better display the size of lacrimal sac, lacrimal mucosal thickness and surrounding soft tissues of lacrimal duct. It is also a good complementary method of lacrimal endoscopy and has guiding significance for individualized treatment in patients with lacrimal duct obstruction.

Organ-specific and Agamous-regulated expression and glycosylation of a pollen tube growth-promoting protein.

Transmitting tissue-specific (TTS) protein is a pollen tube growth-promoting and attracting glycoprotein located in the stylar transmitting tissue extracellular matrix of the pistil of tobacco. The TTS protein backbones have a deduced molecular mass of about 28 kDa, whereas the glycosylated stylar TTS proteins have apparent molecular masses ranging between 50 and 100 kDa. TTS mRNAs and proteins are ectopically produced in transgenic tobacco plants that express either a cauliflower mosaic virus (CaMV) 35S promoter-TTS2 transgene or a CaMV 35S-promoter-NAG1 (NAG1 = Nicotiana tabacum Agamous gene) transgene. However, the patterns of TTS mRNA and protein accumulation and the quality of the TTS proteins produced are different in these two types of transgenic plants. In 35S-TTS transgenic plants, TTS mRNAs and proteins accumulate constitutively in vegetative and floral tissues. However, the ectopically expressed TTS proteins in these transgenic plants accumulate as underglycosylated protein species with apparent molecular masses between 30 and 50 kDa. This indicates that the capacity to produce highly glycosylated TTS proteins is restricted to the stylar transmitting tissue. In 35S-NAG transgenic plants, NAG1 mRNAs accumulate constitutively in vegetative and floral tissues, and TTS mRNAs are induced in the sepals of these plants. Moreover, highly glycosylated TTS proteins in the 50- to 100-kDa molecular mass range accumulate in the sepals of these transgenic, 35S-NAG plants. These results show that the tobacco NAGI gene, together with other yet unidentified regulatory factors, control the expression of TTS genes and the cellular capacity to glycosylate TTS proteins, which are normally expressed very late in the pistil developmental pathway and function in the final stage of floral development. The sepals in the transgenic 35S-NAG plants also support efficient pollen germination and tube growth, similar to what normally occurs in the pistil, and this ability correlates with the accumulation of the highest levels of the 50- to 100-kDa glycosylated TTS proteins.