Objective To investigate the correlation of single nucleotide polymorphisms( SNP) of rs228648 and rs2890565 in urotensin Ⅱ (UTS2)gene with polycystic ovary syndrome(PCOS). Methods The genotypes of two SNPs in UTS2 gene were determined through polymerase chain reaction Tin-shift genotyping method. 101 Chinese Han families trios in Shandong Province consisting of fathers,mothers,and affected daughters with PCOS and 105 healthy women were recruited. The physiological and biochemical parameters including serum follicular stimulating hormone(FSH),luteinizing hormone( LH),testosterone,fasting plasma glucose,and insulin were also measured. Results The distributions of three genotypes and two alleles of SNP rs228648 in UTS2 gene were similar between PCOS group and control group. The distributions of genotypes of SNP rs2890565 in UTS2 gene differed significantly between PCOS group and control group (P<0. 05). The frequency of A allele of SNP rs2890565 in UTS2 gene was significantly higher in PCOS group than that in control group( P<0.05). Transmission disequilibrium test(TDT) did not show significantly biased transmission of two different alleles from parents to affected daughters at rs228648 locus (P>0. 05) ,but showed an excess transmission of A allele from heterozygous parents to affected offspring at rs2890565 locus (P<0. 05). The carriers with GG genotype of SNP rs228648 had significantly higher HOMA-IR compared to those with AA and AG genotypes. The carriers with AA or AG genotype of SNP rs2890565 had significantly higher fasting plasma glucose and fasting insulin compared to those with GG genotypes. HOMA-IR in cases with AA genotype was significantly higher than that with GG genotype. Conclusions There is no association of SNP rs228648 with PCOS, but exists association with insulin resistance. The genetic polymorphism of UTS2 gene rs2890565 may be associated with PCOS,and the higher frequency of A allele is likely to contribute to PCOS.

A sesquiterpene synthase (AsSS4) full-length open reading frame (ORF) cDNA was cloned from wounded stems of Aquilaria sinensis by RT-PCR method. The result showed that the ORF of AsSS4 was 1,698 bp encoding 565 amino acids. Prokaryotic expression vector pET28a-AsSS4 was constructed and transformed into E. coli BL21 (DE3) pLysS. Recombinant AsSS4 protein was obtained after induction by IPTG and SDS-PAGE analysis with a MW of 64 kD. Enzymatic reactions using farnesyl pyrophosphate showed that recombinant AsSS4 protein purified by Ni-agarose gel yielded five sesquiterpene compounds, cyclohexane, 1-ethenyl-1-methyl-2, 4-bis(1-methylethenyl)-, β-elemene, α-guaiene, α-caryophyllene and δ-guaiene. This paper reported the first cloning and functional characterization of AsSS4 gene from A. sinensis, which will establish a foundation for future studies on the molecular mechanisms of wound-induce agarwood formation in A. sinensis

ObjectiveTo compare the effect of aripiprazole and haldol on intelligence and memory in the first-onset schizophrenia patients.MethodsResultsThe total score of PANSS significantly decreased after treatment with aripiprazole and haldol but no significant difference were found between two groups (P>0.05).The total score of WAIS-RC was no significant different between two groups (P>0.05). The total score of WMS was significantly higher in aripiprazole group than in haldol group(P<0.05～0.01).ConclusionAripiprazole is more effective on memory recovery in first-onset schizophrenia than haldol.

Objective:To observe the characteristics of pattern electroretinogram (PERG) and the photopic negative response (PhNR) of flash electroretinogram (FERG) in patients and carriers with Leber hereditary optic neuropathy (LHON).Methods:A cross sectional, observational study. Thirty-two patients (64 eyes) diagnosed with LHON (LHON group) and 15 normal members with the same mutation in patient's family (carrier group) were included in this study from February 2021 to November 2021 in the Department of Ophthalmology of Renmin Hospital of Wuhan University. All patients in LHON group were males (100.0%, 32/32) and the average age was 23.34±7.41 years. In the carrier group of 15 cases (30 eyes), there were 2 males (13.3%, 2/15) and 13 females (86.7%, 12/15). The average age was 43.44±7.65 years. Twenty-four healthy subjects (48 eyes) in the same period were selected as the control group. Among them, there were 8 males (33.3%, 8/24) and 16 females (66.7%, 16/24). The average age was 23.42±2.54 years. All subjects were examined with the GT-2008V-VI visual electrophysiology instrument of Chongqing Gotec Medical Equipment Limited Company for PERG and FERG. P50 and N95 amplitudes of PERG and PhNR, a wave and b wave amplitudes of FERG were recorded. The peripapillary retinal nerve fiber layer (pRNFL) thicknesses of the nasal, superior, temporal, inferior and average quadrants were measured by spectral domain optical coherence tomography (SD-OCT). The amplitudes of a wave, b wave, PhNR, P50 wave, N95 wave and pRNFL thickness between the three groups were compared by one-way ANOVA. Pearson correlation analysis was used to analyze the correlation between different parameters.Results:Compared with the control group, the amplitudes of PhNR in LHON group and carrier group decreased significantly ( F=11.973, P<0.001). The results of correlation analysis showed that the amplitude of PhNR in LHON group was significantly correlated with the thickness of nasal and temporal pRNFL ( r=0.249, 0.272; P=0.048, 0.030). There was no significant difference in P50 wave amplitude between patients, carriers and controls ( F=1.342, P=0.265). There was no significant difference in N95 wave amplitude between patients and controls ( P=0.960). Conclusion:The PhNR amplitudes of FERG in LHON patients and carriers decrease significantly compared to controls.

Objective To investigate the correlation of substantia nigra hyperecho with essential tremor (ET) and Parkinson disease (PD).Methods The clinical data of 158 patients with ET or PD who underwent transcranial ultrasonography in Tongji Hospital from March 2016 to March 2018 were retrospectively analyzed.There were 35 patients with ET (ET group),113 patients with PD who had no previous history of ET (PD group),and 10 PD patients with previous history of ET (ET-PD group).And 58 healthy subjects served as controls (control group).The hyperechoic area of substantia nigra in different groups was compared.Results The hyperechoic areas of the substantia nigra were [0 (0,0)]cm2 (control group),[0.27(0,0.41)]cm2 (ET group),[0.33(0.21,0.40)]cm2 (ET-PD group) and [0.35(0.29,0.45)]cm2 (PD group);the differences between control group and ET group,between the ET group and PD group were statistically significant (Z=-5.24,P=0.01;Z=-3.09,P=0.02),and there were no significant difference between the ET group and ET-PD group,between ET-PD group and PD group (Z=-0.98,P=0.32;t=-0.98,P=0.33).The ratio of substantia nigra hyperechoic positive to negative in ET-PD group was 9.00 (9/1),while that in ET group was 0.94 (17/18) (OR=9.53,95％ CI:1.09-83.43,x2=3.91,P=0.04).Conclusion Substantia nigra hyperecho is an objective imaging indicator for patients with ET and PD,and has a certain differential value for their diagnosis.

Objective:To explore the relationship between the changes of midbrain raphe nucleus echo and blood trace metals, non-motor symptoms in patients with Parkinson′s disease (PD).Methods:A total of 177 patients with PD were recruited from Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology from November 2017 to December 2019. Their non-motor symptoms were assessed by a series of PD-related scales. All subjects completed transcranial color-coded sonography, and were divided into two groups according to the continuity of raphe nucleus echo. The difference of trace metals in peripheral blood and the scores of non-motor symptoms were analyzed.Results:The serum iron level of PD patients in the continuous echo of raphe nucleus group (117 cases; 15.33 （11.30, 18.93） μmol/L) was higher than that of the discontinuous group (60 cases; 12.52 （4.15, 16.00） μmol/L, t=-2.294, P=0.022), so was the scores of Scale for outcomes in PD for Autonomic Symptoms (34.00 （28.00, 39.00） vs31.00 （26.25, 36.00）， t=-2.044, P=0.041). There was no significant difference in the level of serum copper, hemoglobin, ceruloplasmin, and the scores of Movement Disorder Society Unified Parkinson′s Disease Rating Scale, Parkinson′s Disease Sleeping Scale, 39 items Parkinson′s Disease Questionnaire, Hamilton Depression Rating Scale and Hamilton Anxiety Rating Scale between the two groups. Conclusion:There was significant difference in serum iron levels and the scores of SCOPA-AUT between the two groups, but no difference in depression, sleep, and daily living ability.

Objective:To apply digital analysis to quantify hyperechogenicity of substantia nigra, and explore its clinical value for diagnosis of Parkinson′s disease (PD).Methods:The cross-sectional study included 652 PD patients (PD group) and 99 healthy controls (healthy control group) from November 2017 to October 2020 in Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology. All subjects underwent transcranial sonography. The diagnostic accuracy of substantia nigra hyperechogenicity using digital analysis was compared with that in a manual measurement in PD. Furthermore, the receiver operating characteristic (ROC) curve analysis was applied to explore its diagnosis value in PD.Results:There were 482 subjects including 400 in the PD group and 82 in the healthy control group, whose quantified results of substantia nigra hyperechogenicity could be used for analysis. The ROC analysis showed that the area under the curve of the quantified larger substantia nigra hyperechoic region detection for diagnosing PD was 0.858 (95% CI 0.805-0.910), the sensitivity was 87.8%, and the specificity was 73.2%, consistent with that of doctors (area under the curve: 0.884). Further more, among these PD patients, there was no correlation between larger substantia nigra hyperechogenicity and age, age of onset, course of disease, non-motor symptoms, and motor symptoms (all P>0.05). Conclusions:Digital analysis was used to quantify the changes in substantia nigra hyperechogenicity in this seudy. The results showed that diagnostic accuracy for PD based on digital analysis was consistent with that of experienced clinicians.

OBJECTIVETo characterize the mutation spectrum of phenylalanine hydroxylase (PAH) gene and perform prenatal diagnosis for families with classical phenylketonuria.

METHODSBy stratified sequencing, mutations were detected in the exons and flaking introns of PAH gene of 44 families with classical phenylketonuria. 47 fetuses were diagnosed by combined sequencing with linkage analysis of three common short tandem repeats (STR) (PAH-STR, PAH-26 and PAH-32) in the PAH gene.

RESULTSThirty-one types of mutations were identified. A total of 84 mutations were identified in 88 alleles (95.45%), in which the most common mutation have been R243Q (21.59%), EX6-96A>G (6.82%), IVS4-1G>A (5.86%) and IVS7+2T>A (5.86%). Most mutations were found in exons 3, 5, 6, 7, 11 and 12. The polymorphism information content (PIC) of these three STR markers was 0.71 (PAH-STR), 0.48 (PAH-26) and 0.40 (PAH-32), respectively. Prenatal diagnosis was performed successfully with the combined method in 47 fetuses of 44 classical phenylketonuria families. Among them, 11 (23.4%) were diagnosed as affected, 24 (51.1%) as carriers, and 12 (25.5%) as unaffected.

CONCLUSIONPrenatal diagnosis can be achieved efficiently and accurately by stratified sequencing of PAH gene and linkage analysis of STR for classical phenylketonuria families.

Adolescent ; Adult ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Fetal Diseases ; diagnosis ; enzymology ; genetics ; Genetic Testing ; Humans ; Infant ; Infant, Newborn ; Male ; Microsatellite Repeats ; Middle Aged ; Pedigree ; Phenylalanine Hydroxylase ; genetics ; Phenylketonurias ; diagnosis ; enzymology ; genetics ; Point Mutation ; Pregnancy ; Prenatal Diagnosis ; Young Adult

Objective:To investigate the clinical value of isolated fetal echogenic bowel (FEB) as an indicator for invasive prenatal diagnosis.Methods:This retrospective study enrolled 183 pregnant women who were diagnosed with isolated FEB and underwent invasive prenatal diagnosis in Fujian Maternity and Child Health Hospital from August 2013 to January 2021. Clinical data including the results of conventional karyotyping and chromosomal microarray analysis (CMA), cytomegalovirus (CMV) DNA loads in amniotic fluid and pregnancy outcomes were reviewed analyzed. Chi-square test was used for statistical analysis Results:Karyotyping was performed on all of the 183 fetuses and three (1.64%) aneuploidies (one case of trisomy 21, one trisomy 18 and one 47,XYY syndrome) were detected. One trisomy 21 and four pathogenic (P)/likely pathogenic (LP) copy number variation (CNV) were detected among 108 fetuses who received CMA. The detection rate of P/LP chromosomal abnormalities by CMA was higher than that by karyotyping, but there was no significant difference between them [4.63% (5/108) vs 0.93% (1/108), χ 2=1.54, P>0.05]. In addition, three cases of variants of uncertain significance (VOUS) were detected by CMA. CMV DNA loads of fetal cells in the amniotic fluid samples of the 166 cases were determined, and only one (0.6%) was positive (CMV DNA up to 7.01×10 6 copies/ml), and no abnormalities were found in karyotype analysis and CMA detection. A total of 176 cases were followed up, and among them only one case of intrauterine infection and seven cases (three aneuploidies and four P/LP CNV) of chromosomal abnormalities were terminated after genetic counseling. Three fetuses with VOUS and other 165 fetuses without chromosomal abnormalities had a good prognosis after birth. Conclusions:Isolated FEB may be the abnormal ultrasound finding in fetuses with chromosomal abnormalities or CMV infection. Prenatal genetic testing and the exclusion of intrauterine infection are important for management during pregnancy and prognosis assessment of FEB.

Objective:To compare the optimal conditions, virus yield, viral titer and cell metabolism between culturing influenza virus H1N1 vaccine strain in MDCK and MDCK-G1 cells.Methods:The optimal culture conditions were investigated using chessboard method. The hemagglutination titer, half of the tissue infection dose (TCID 50) and the metabolism of glucose and lactic acid were monitored and compared between the two cell lines. Results:After MDCK-G1 cells were inoculated with H1N1 at the multiplicity of infection (MOI) of 0.001 with the presence of 1 μg/ml of trypsin, the hemagglutination titer reached the peak of 1∶512 at 72 h and the viral titer was 10 7.4TCID 50/ml. In the MDCK cell line group, the hemagglutination titer reached the peak of 1∶256 at 72 h and the viral titer was 10 6.6TCID 50/ml when using H1N1 at MOI=0.0001 and 1 μg/ml of trypsin. Conclusions:MDCK-G1 cells were more suitable than MDCK cells for the proliferation of influenza virus. This study provided reference data for further research on cell-derived influenza vaccine.