Objective To investigate mutation spectrums of α- and β-haemoglobin genes in thalassemia patients and carriers in Yunnan province,and to establish procedures on prenatal gene diagnosis.MethodsTotally 10 033 counseling couples and pregnant women,and 22 cases of children with moderate or severe thalassemia were recruited from 5 parts of Yunnan Province,middle,western,eastern,southern and northern areas, during July 2009 to July 2011.Medical records, including results of haemoglobin electrophoresis,blood routine examination,and gene diagnosis of subjects were collected and saved in an database in Excel software by the Key Laboratory for Birth Defects and Genetic Diseases.Using multiple gap-PCR and PCR-reversed dot blotting kits, DNA samples collected from 1077 cases of haematological positive thalassemia patients and carriers were tested to determine common mutations of the α-or β-haemoglobin genes.The codon regions of haemoglobin genes were sequenced by the Sanger sequencing in cases that the mutation tests were negative.Mutation spectrums of α- and β-haemoglobin genes were concluded.Prenatal gene diagnosis was offered to fetuses who had risk of thalassemia major.Results( 1 ) In 1077 cases of haemological screen positive subjects,deletions and mutations of α-haemoglobin gene were tested in 119 subjects among 347 cases suspected as α-thalassemia patients and carriers.Five kinds of deletions and mutations on α-haemoglobin gene were found.In 104 subjects,four kinds of common deletions and mutations onα-haemoglobin gene were determined:--SEA, -α3.7, αCS α,-α4.2.Other 14 subjects were double heterozygotes with haemoglobin H disease and severe α-thalassemia phenotypes.A rare mutation of insertion and deletion in α2 haemoglobin gene intron,α301-24-301-23 indel,was found in one carrier subject.(2)In 1077 cases of haemological screen positive subjects,deletions and mutations of β-haemoglobin gene were tested in 297 subjects among 730 cases suspected as β-thalassemia patients and carriers.Sixteen kinds of β-haemoglobin gene mutations were found,including 7 cases of rare abnormal haemoglobinopathy patients with β-haemoglobin gene mutations.In one case with β + phenotype patient,the Codon 5 (-CT)mutation at β-haemoglobin gene was found (firstly reported in China ). (3) Three fetuses with high riskS of α-thalassemia were accepted for prenatal diagnosis.One case of Hb Bart's hydrops syndrome fetus with the genotype --SEA/--SEA,and one case of mild α-thalassemia fetus with the genotype αCS α/αα were found.Another one fetus was found with normal α-haemoglobin.In 6 fetuses accepted for prenatal diagnosis due to high risks of β-thalassemia,one case of β-thalassemia major with the genotype CD17( A→T)/-28 (A→G) was found,3 fetuses were heterozygote carriers,and 2 fetuses had normal genotypes without mutations found in their parents.Medical terminations for 2 fetuses with severe thalassemia were made according to the choice of pregnant women.Other 7 pregnancies continued to term.Anemia or growth retardation was not found in the 7 infants when following up after given-birth 6 to 12 months.Conclusions The mutation spectrums of α- and β-haemoglobin genes of thalassemia patients and αarriers.in Yunnan province are special,in which β-haemoglobin gene exits more polymorphism in the mutation spectrum.Carrier screening in pregnant women,and offering prenatal gene diagnosis to the high risk pregnancies should be an efficient strategy to prevent thalassemia major.

Objective To discuss the diagnostic value of ultrasonography in piriformis syndrome . Methods Ultrasonography was performed in thirty‐eight patients with unilateral piriformis syndrome and forty healthy volunteers . The morphological structures and the internal echoes of their bilateral piriformises and sciatic nerves were observed and their thicknesses were measured . These parameters of the patients and voluteers were recorded and compared . Results The ultrasonographic images of piriformis and sciatic nerve of the healthy voluteers showed no abnormal change . The thickness difference of their bilateral piriformises and sciatic nerves had no statistical significance ( P > 0 .05 ) . The ultrasonography image of the morphological structure and the internal echo of the sick side piriformis and sciatic nerve of the patients with piriformis syndrome showed a change ,that the sick side piriformis was significantly thicker than the healthy side piriformis [(25 .74 ± 3 .12) mm vs (22 .48 ± 2 .60) mm , P < 0 .05] . The area under the operator characteristic curve ( AUC ) for the thickness difference of bilateral piriformises in diagnosing piriformis syndrome was 0 .896 ,with the optimal cut‐off value of 2 .15 mm . However ,the thickness difference of their bilateral sciatic nerves had no statistical significance ( P >0 .05) . Conclusions Ultrasonography can show piriformis and sciatic nerve clearly . The ultrasonographic images and the thickness difference of the bilateral piriformises is helpful to diagnose piriformis syndrome ,and can provide more informations for clinic .

Objective To explore the clinical value of genetic diagnosis of SMA,the homozygous deletion of survival motor neuron 1 (SMN1) gene in suspected spinal muscular atrophy (SMA) patients were analyzed in this study.Methods A total of 154 patients suspected with SMA and 20 healthy volunteers were recruited from January 2007 to December 2014 in the Genetic Diagnosis Center of the First People's Hospital of Yunnan Province and the Department of Neurology of the Fourth Affiliated Hospital of Kunming Medical University.Potential deletions in exons 7 and 8 of SMN1 gene were screened by use of polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) method in both 154 patients suspected with SMA and 20 healthy volunteers.The frequencies of the deletions in exons 7 and 8 of SMN1 were calculated and statistical analysis of different deletion types among 3 SMA groups was performed with SPSS 13.0 software package.Results Among 154 suspected SMA patients,101 cases with homozygous deletions of exon 7 of SMN1 gene were detected,which accounted 65.6％ (101/154) of the suspected SMA patients.Among the 101 SMA patients,97.0％ (98/101) of the patients with both homozygous deletions of exons 7 and 8 for SMN1 gene and 3.0％ (3/101) of the patients with homozygous deletions of only exon 7 for SMN1 gene were detected.The patient with only deletion of exon 8 for SMN1 gene was notdetected.Four cases with negative results were subjected to be followed-up,but they were characteristic of SMA symptom by clinical re-visit.Thus,total 105 patients were confirmed with SMA,among them,68 were type Ⅰ SMA,27 were type Ⅱ SMA,and 10 were type Ⅲl SMA,which accounted for 64.8％ (68/105),25.7％ (27/105) and 9.5％ (10/105) of the SMA patients,respectively.Type Ⅳ SMA was not observed in these patients.No deletion was detected among 20 healthy volunteers.Conclusions PCR-RFLP assay is a noninvasive,simple,high sensitive and specific method for SMA diagnosis,which can be considered as the first-line genetic test for the suspected SMA patients.It will help to improve the accuracy of clinical diagnosis and the detection rate by strengthening the clinical diagnostic criteria and re-evaluating the suspected patients after negative genetic diagnosis.

Objective To explore the relations of clinical phenotypes of type Ⅰ-ⅣV spinal muscular atrophy (SMA) with neural electrophysiological features and survival motor neuron (SMN)gene.Methods A total of 85 patients with SMA,including 46 with infantile form in which 19 of type Ⅰ and 27 of type Ⅱ,24 with juvenile form (type Ⅲ),and 15 with adult form (type ⅣV),were involved in this clinical study.Their clinical data were analyzed.The neural conduction,needle electromyography (EMG)and SMN1 gene deletion were analyzed.Results There existed different clinical features among patients who suffered from type Ⅰ to type ⅣV SMA.However,the major clinical features of SMA were displayed by progressively aggravating of flaccid paralysis in muscles of the four limbs,and the younger of the patients,the more serious of the clinical manifestations.EMG exhibited neurogenic lesion in all 43 SMA patients,33 patients presented generalized neurogenic lesions,and the abnormal degree of muscles in lower limbs was more severe than that of upper limbs,and the proximal muscles was more severe than that of the distal ones.The abnormal rate of spontaneous potential,weak contraction with raise potential and amplitude of compound motor active potential in adult and juvenile SMA were significantly lower than those in infantile SMA.SMN1 gene exon 7 and 8 were detected in all 85 patients with SMA.A total of 61 patients were found with deletion of exon 7 and/or 8 in SMN1 gene.Infantile SMA patients enjoyed 95.7％ (44/46) detection rate,juvenile SMA patients enjoyed 70.8％ (17/24) detection rate;no adult SMA patients were found with deletion ofexon 7 and/or 8 in SMN1 gene.Conclusions The more serious of clinical manifestations in SMA patients,the higher abnormality rote in electrophysiological tests.The exons deletion in SMN1 gene could result in alterations of SMA phenotypes,but it has nothing to do with the severity of SMA.Gene deletion analysis of SMN1 gene can be considered as the preferred fimal diagnosis method for infantile SMA patients.But as for juvenile form,its diagnosis depending on gene deletion analysis of SMN1 gene will be analyzed with precaution.While as for adult form of SMA,the incidence of SMA may be independent with SMN1 gene deletion.Therefore,generally,SMN1 gene assay is not taken as the routine diagnose method for adult form SMA.

Objective:Patients with acute exacerbation of chronic obstructive pulmonary disease(AECOPD)combined with severe type Ⅱ respiratory failure have a high probability of ventilation failure using conventional non-invasive positive pressure ventilation(NPPV).This study aims to investigate the clinical efficacy of high intensity NPPV(HI-NPPV)for the treatment of AECOPD combined with severe type Ⅱ respiratory failure. Methods:The data of patients with AECOPD combined with severe type Ⅱ respiratory failure(blood gas analysis pH≤7.25)treated with NPPV in the Second Affiliated Hospital of Chongqing Medical University from July 2013 to July 2023 were collected to conduct a retrospective case-control study.The patients were divided into 2 groups according to the inspired positive airway pressure(IPAP)used during the NPPV treatment:a NPPV group(IPAP<20 cmH2O,1 cmH2O=0.098 kPa)and a HI-NPPV group(20 cmH2O≤IPAP<30 cmH2O).Ninety-nine and 95 patients were included in the NPPV group and the HI-NPPV group,respectively.A total of 86 pairs of data were matched using propensity score matching(PSM)for data matching.The primary outcome indexes(mortality and tracheal intubation rate)and secondary outcome indexes[blood gas analysis pH,arterial partial pressure of oxygen(PaO2)and arterial partial pressure of carbon dioxide(PaCO2),adverse reaction rate,and length of hospitalization]were compared between the 2 groups. Results:The tracheal intubation rates of the NPPV group and the HI-NPPV group were 6.98％and 1.16％,respectively,and the difference between the 2 groups was statistically significant(χ2=4.32,P<0.05);the mortality of the NPPV group and the HI-NPPV group was 23.26％and 9.30％,respectively,and the difference between the 2 groups was statistically significant(χ2=11.64,P<0.01).The PaO2 at 24 h and 48 h after treatment of the HI-NPPV group was higher than that of the NPPV group,and the PaCO2 of the HI-NPPV group was lower than that of the NPPV group,and the differences were statistically significant(all P<0.05).The differences of pH at 24 h and 48 h after treatment between the 2 groups were not statistically significant(both P>0.05).The differences between the 2 groups in adverse reaction rate and hospitalization length were not statistically significant(both P>0.05). Conclusion:HI-NPPV can reduce mortality and tracheal intubation rates by rapidly improving the ventilation of patients with AECOPD combined with severe type Ⅱ respiratory failure.This study provides a new idea for the treatment of patients with AECOPD combined with severe type Ⅱ respiratory failure.

Objective:To analyze the genetic mutation characteristics of glucose-6-phosphate dehydrogenase (G6PD) deficiency among infants in Kunming.Methods:A total of 15 533 infants (7 994 males and 7 539 females) born in Kunming from January 1, 2018, to December 31, 2020, with an age range of 2 to 44 days, were selected. G6PD enzyme activity and gene mutation types were detected using fluorescence quantitative analysis, multicolor melting curve analysis (MMCA), and Sanger sequencing. Droplet digital PCR (ddPCR) was used for quantitative analysis of a newly identified variant family to determine the mutant allele proportion in family members. Meanwhile,the protein structure model and pathogenicity prediction of the novel variant were analyzed.Data analysis was conducted using SPSS 26.0. Specifically, chi-square tests were used for the detection rates of G6PD enzyme activity and gene mutations between different genders. One-way analysis of variance (ANOVA) was used for the comparison of enzyme activity among different mutation types.Results:Among 15 533 infants, 143 cases (129 males and 14 females) were tested positive for G6PD activity, with a detection rate of 0.92% (143/15 533). The difference in detection rates between males and females was statistically significant (χ 2=96.76, P<0.001). Out of 89 enzyme activity-positive cases (83 males and 6 females) underwent genetic testing, 77 (72 males and 5 females) were detected by MMCAand other 12 negative samples were underwent further Sanger sequencing, revealing mutations in 6 samples, all of which were males. Among the 83 individuals with gene mutations, 78 had heterozygous mutations, 1 had a homozygous mutation, and 4 had compound heterozygous mutations. A total of 12 mutation types were detected, with G6PD c.487G>A, c.1024C>T, c.1388G>A, and c.1376G>T being the most common, accounting for 74.70% (62/83) of all mutation types. The average G6PD enzyme activity of c.1376G>T was the lowest, and the differences were statistically significant compared to the average enzyme activity of the other three mutations ( P<0.05). One male infant with a newly identified G6PD c.242G>C mutation was detected, predicted to be pathogenic. ddPCR confirmed that the mother of the affected child was a c.242G>C mutant chimera, with a chimera proportion of 6.66%. Conclusions:In the Kunming region, the predominant G6PD deficiency gene mutation is c.487G>A, with the detection of a novel G6PD c.242G>C mutation. The application of ddPCR technology can assist in detecting the proportion of mutation chimeras.

OBJECTIVETo provide genetic analysis for a pregnant woman with chromosomal translocations and intellectual disability, and to provide prenatal diagnosis for her fetus.

METHODSRoutine G-banding was performed to analyze the karyotypes of the woman and her fetus. Copy number variants were determined with array comparative genomic hybridization (array-CGH).

RESULTSThe pregnant woman has carried an apparently balanced translocation involving chromosomes 1, 2, 6 and 7, with a karyotype of 46, XX, t(1;2) (p22;p23), t(6;7) (q21;p15). The karyotype of her fetus was ascertained as 46, XY, t(6;7) (q21;p15) mat. Array-CGH has detected a 4 Mb microdeletion at 6q22.1-q22.31 (115 311 507-119 332 956) in both individuals. As the 6q22.1-q22.31 microdeletion may be associated with the main clinical manifestations of the woman, the family decided to terminate the pregnancy. The fetus was male and appeared to have no obvious abnormality.

CONCLUSIONPrenatal diagnosis for pregnant women with translocations and mental retardation is a challenging task. Combined application of cytogenetic analysis and array-CGH may facilitate the diagnosis and genetic counseling.

Adult ; Female ; Fetus ; abnormalities ; Genetic Testing ; methods ; Humans ; Intellectual Disability ; genetics ; Male ; Pregnancy ; Prenatal Diagnosis ; methods ; Translocation, Genetic ; genetics ; Young Adult

OBJECTIVE: To perform carrier screening for spinal muscular atrophy (SMA) among 3049 reproductive-age individuals from Yunnan region and determine the copy number of survival motor neuron (SMN) gene and carrier frequencies. METHODS: Multiplex ligation-dependent probe amplification (MLPA) was used to determine the copy number of exon 7 of SMN1 and SMN2 genes and identify those with a single copy of SMN1 gene. Prenatal diagnosis was performed for couples whom were both found to be SMA carriers. RESULTS: In total 62 SMA carriers were identified among the 3049 subjects, which yielded a carrier frequency of 1 in 49 (2.03%). No statistical difference was found in the carrier frequency between males and females (1.91% vs. 2.30%, P>0.05). Respectively, 1.3% (41/3049) and 0.69% (21/3049) of the carriers were caused by heterozygous deletion and conversion of the SMN1 gene. The average copy number for SMN1 alleles was 1.99. Two couples were found to be both as SMA carriers, for whom the birth of an affected fetus was avoided by prenatal diagnosis. CONCLUSION No difference was found in the carrier frequency of SMA-related mutations between the two genders in Yunnan region, which was in keeping to an autosomal recessive inheritance pattern. Determination of the carrier frequency for SMA and SMN gene variants may provide a basis for genetic counseling and prenatal diagnosis for the disease.
China ; Female ; Genetic Carrier Screening ; Genetic Counseling ; Genetic Variation ; Heterozygote ; Humans ; Male ; Muscular Atrophy, Spinal ; genetics ; Pregnancy ; Prenatal Diagnosis ; Survival of Motor Neuron 1 Protein ; genetics ; Survival of Motor Neuron 2 Protein ; genetics

Objective:To investigate the clinical features and associated chronic pain in corticobasal syndrome (CBS).Methods:Clinical data of 8 patients diagnosed as probable CBS or possible CBS admitted to Beijing Hospital during January 2010 to June 2020 were retrospectively analyzed. The clinical information included sex, age, course of disease, chief complaint, neurological examination, blood biochemistry, tumor marker, infection and other laboratory tests; the neuropsychological evaluation included Mini-Mental State Examination (MMSE) scale and Hamilton Depression Scale (HAMD); the imaging studies included cranial magnetic resonance imaging (MRI) and/or 18F-Fluorodeoxyglucose positron emission tomography ( 18F-FDG PET). Results:The main clinical manifestations were asymmetrical movement disorders, including rigidity, tremor, myoclonus and abnormalities in posture and gait. Patients showed poor response to levodopa treatment. Among 8 patients, 7 had apraxia, 5 patients had alien hand, and 5 patients had various degrees of cognitive dysfunction. The cranial MRI demonstrated mild cerebral atrophy which was slightly more severe in the contralateral side of the initially affected limb in 7 of the 8 patients. The 18F-FDG PET scan revealed asymmetric decreased metabolism in the frontal, parietal, temporal, and occipital lobe, as well as in basal ganglia, which was more severe in the contralateral side of the initially affected limb in 5 of the 8 patients. Six of the 8 patients were associated with pain, including dystonic pain in 3 patients, neuropathic pain in 1 patient, musculoskeletal pain in 1 patient, and unexplained pain in 1 patient. Pain was the onset symptom in 1 patient and pain was relieved by taking levodopa in another patient. Conclusions:CBS is characterized by asymmetric dyskinesia and cognitive impairment, and often associated with apraxia, cortical sensory deficits, and alien limb. The MRI and PET are helpful for CBS diagnosis. Pain may be one of the common non-motor symptoms in CBS.