To further uncover the scientific significance and molecular mechanism of the Chinese herbs with pungent hot or warm natures, endogenous and exogenous expression systems were established by isolation of dorsal root ganglion (DRG) neurons and transfection of HEK293 cells with TRPV1 channel gene separately. On this basis, the regulation action of capsaicin, one main ingredient from chili pepper, on TRPV1 channel was further explored by using confocal microscope. Besides, the three-sites one-unit technique and method were constructed based on the brown adipose tissue (BAT), anal and tail skin temperatures. Then the effect of capsaicin on mouse energy metabolism was evaluated. Both endogenous and exogenous TRPV1 channel could be activated and this action could be specifically blocked by the TRPV1 channel inhibitor capsazepine. Simultaneously, the mice's core body temperature and BAT temperature fall down and then go up, accompanied by the increase of temperature of the mice's tail skin. Promotion of the energy metabolism by activation of TRPV1 channel might be the common way for the pungent-hot (warm) herbs to demonstrate their natures.

Aim The TRPV1 plasmid was transiently transfected into human embryonic kidney HEK 293T cells to establish the heterologous expression system of TRPV1-channel. Methods The transfection efficiency was confirmed under fluorescence mi-croscope and the TRPV1 protein expression was identified by u-sing Western blot, and the functional characteristics of the chan-nel were studied by using the method of confocal microscopy. Results The transfection rate could reach 40% ~50%; the transfected cells were found to have a clear band at the corre-sponding position that TRPV1 should be, which indicated that TRPV1 channel protein was expressed in the transfected cells. The confocal microscopy imaging result showed that the trans-fected HEK 293T cells were activated by TRPV1 channel ago-nist. Conclusion Transient transfection of HEK 293T cells with TRPV1 channel is successfully constructed and the heterol-ogous TRPV1 channel is verified to have normal calcium-media-ting function.

Objective To investigate the value of array-based comparative genomic hybridization (array-CGH) technique for the detection of chromosomal analysis of miscarried embryo, and to provide genetic counseling for couples with spontaneous abortion. Methods Totally 382 patients who underwent miscarriage were enrolled in this study. All aborted tissues were analyzed with conventional cytogenetic karyotyping and array-CGH, respectively. Results Through genetic analysis, all of the 382 specimens were successfully analyzed by array-CGH (100.0%, 382/382), and the detection rate of chromosomal aberrations was 46.6% (178/382). However, conventional karyotype analysis was successfully performed in 281 cases (73.6%, 281/382), and 113 (40.2%, 113/281) were found with chromosomal aberrations. Of these 178 samples identified by array-CGH, 163 samples (91.6%, 163/178) were aneuploidy, 15 samples (8.4%, 15/178) were segmental deletion and (or) duplication cases. Four of 10 cases with small segmental deletion and duplication were validated to be transferred from their fathers or mathers who were carriers of submicroscopic reciprocal translocation. Of these 113 abnormal karyotypes founded by conventional karyotyping, 108 cases (95.6%, 108/113) were aneuploidy and 5 cases (4.4%, 5/113) had chromosome structural aberrations. Most array-CGH results were consistent with conventional karyotyping but with 3 cases of discrepancy, which included 2 cases of triploids, 1 case of low-level mosaicism that undetcted by array-CGH. Conclusions Compared with conventional karyotyping, there is an increased detection rate of chromosomal abnormalities when array-CGH is used to analyse the products of conception, primarilly because of its sucess with nonviable tissues. It could be a first-line method to determine the reason of miscarrage with higher accuracy and sensitivity.

Objective:To correlate multiphase CT angiography (mCTA), serum lipid peroxidation (LPO) and thrombus precursor protein (TpP) levels with recurrence of acute cerebral infarction (ACI) in older adults and investigate the value of these indicators in the predication of ACI recurrence.Methods:A total of 128 older adult patients with ACI who received treatment in Ningbo Medical Center Lihuili Hospital, China between January 2019 and January 2020 were included in this study. All of them were followed up for 1 year. They were divided into ACI recurrence group ( n = 29) and no ACI recurrence group ( n = 99) according to whether they had recurrent cerebral infarction. All patients underwent mCTA. Maas system and Tan score were used according to mCTA images. Serum TpP level was measured using enzyme-linked immunosorbent assay. Serum LPO level was measured using Yagi's fluorescence method. Multiple linear regression analysis was used for correlation analysis. The receiver operating characteristic (ROC) curve was used to evaluate the efficacy of mCTA and serum LPO and TpP levels in the diagnosis of ACI. Results:Tan score in the ACI recurrence group was significantly lower than that in the no ACI recurrence group [(1.06 ± 0.26) points vs. (1.89 ± 0.82) points, t = 5.35, P < 0.05]. Serum TpP and LPO levels in the ACI recurrence group were (7.22 ± 1.35) mmol/L and (11.23 ± 2.58) nmol/mL, respectively, which were significantly higher than those in the no ACI recurrence group [(3.06 ± 0.28) mmol/L, (7.23 ± 0.37) nmol/mL, t = 28.86, 15.04, both P < 0.001]. ACI recurrence in older adult patients was correlated with Tan score and serum LPO and TpP levels (both P < 0.05). The sensitivity of mCTA combined with serum LPO and TpP levels in the diagnosis of ACI in older adults was 93.10%-96.60% and its specificity was 100.00%. The ROC curve analysis showed that the area under the ROC of mCTA, LPO and TpP in the prediction of ACI recurrence in older adults was 0.986 (95% CI = 0.966-1.000), 0.976 (95% CI = 0.930-1.000) and 0.968 (95% CI = 0.905-1.000), respectively. Conclusion:ACI recurrence in older adults is correlated with Tan score and serum LPO and TpP levels. mCTA, Tan score, and serum LPO and TpP levels have high sensitivity and specificity in the diagnosis of ACI recurrence in older adults, and therefore have a high diagnostic value.

OBJECTIVETo study the genetic polymorphisms and mutations of 20 frequently used autosomal microsatellites among ethnic Hans from Henan.

METHODSPeripheral blood samples of 2604 individuals were collected. DNA was amplified and genotyped using a PowerPlex(TM) 21 system. The frequencies, forensic parameters and mutation rates of the 20 short tandem repeat (STR) loci were analyzed.

RESULTSA total of 323 alleles were found in this population and the allelic frequencies have ranged from 0.0003 to 0.5144. Except for D3S1358, TH01 and TPOX, mutations have been found in all of the remaining 17 STR loci, which totaled 47, with mutation rates ranging from 0 to 3.46 × 10.

CONCLUSIONThe 20 STR loci selected by the PowerPlex(TM) 21 system are highly polymorphic among ethnic Hans from Henan, and may be of great value in forensic and human population studies. As no similar study has been carried out previously, above results may be of great value for individual discrimination and paternal testing.

Adolescent ; Adult ; Alleles ; Asian Continental Ancestry Group ; ethnology ; genetics ; Child ; Child, Preschool ; China ; ethnology ; Female ; Genotype ; Humans ; Male ; Microsatellite Repeats ; Mutation ; Pedigree ; Polymorphism, Genetic ; Young Adult

OBJECTIVETo analyze a child with facial abnormalities with combined cytogenetic and molecular techniques and delineate its clinical phenotype.

METHODSNeuropsychological profile of the child was analyzed. Color Doppler, CT and MRI were used for detecting the nodules in the body. Conventional peripheral blood karyotypes of the child and his parents were analyzed with G-banding. Array-comparative genomic hybridization (aCGH) was performed to detect minor structural chromosomal abnormalities.

RESULTSThe child had mental retardation, maxillofacial dysmorphism on the right side, and irregular solid nodules on the back. The karyotypes of the child and his parents were all normal, while aCGH has identified a de novo constitutive 1.2 Mb deletion at 17q11.2 in the child. The aCGH results of his parents were normal.

CONCLUSIONThe de novo 17q11.2 microdeletion probably underlies the facial abnormalities and neurofibromatosis in the patient.

Child, Preschool ; Chromosome Banding ; Chromosome Deletion ; Chromosomes, Human, Pair 17 ; genetics ; Comparative Genomic Hybridization ; Humans ; Intellectual Disability ; genetics ; Karyotyping ; Male ; Maxillofacial Abnormalities ; genetics ; Phenotype ; Smith-Magenis Syndrome ; genetics

Objective To explore the role of next-generation sequencing(NGS)technology in the assisted diagnosis of RA-Sopathies.Methods Peripheral blood was extracted from 1 child patient with suspected Noonan syndrome and her parents,and the gene mutations were detected by adopting the aCGH and NGS.The results were verified by Sanger sequencing.Results The NGS results revealed that the heterozygous mutation of c.1406G>A existed in BRAF gene,and the results of Sanger sequencing in this child case was consistent with the NGS results.The Sanger sequencing results in her parents showed the locus was G/G wild type. Conclusion This child case was diagnosed as CFC.NGS plays a good auxiliary role in the differentiation diagnosis of RASopathies.

OBJECTIVE: To explore the molecular mechanism of a girl with developmental delay and intellectual disability. METHODS: Chromosomal karotypes of the child and her parents were analyzed with routine G-banding method. Their genomic DNA was also analyzed with array comparative genomic hybridization (aCGH) for chromosomal duplications/deletions. RESULTS: No karyotypic abnormality was detected in the child and her parents, while aCGH has identified a de novo 3.37 Mb deletion at 17p11.2 in the child. CONCLUSION The child was diagnosed with Smith-Magenis syndrome, for which RAI1 may be the causative gene.
Child ; Chromosome Deletion ; Chromosome Duplication ; Chromosomes, Human, Pair 17 ; genetics ; Comparative Genomic Hybridization ; Female ; Humans ; Karyotyping ; Smith-Magenis Syndrome ; genetics