Establishment of human-induced pluripotent stem cell GZHMCi0011-A from peripheral blood mononuclear cells from a volunteer with 14/63 CAG repeats of the ATXN3 mutation.

Spinocerebellar ataxia type 3 (SCA3) is a genetic degeneration disease of the nervous system with ataxia as the main clinical manifestation, and the most frequent subtype of SCA3 is known to be caused by CAG repeat expansions of more than 55 units in ATXN3. In this study, we used peripheral blood mononuclear cells (PBMCs) from a volunteer with 14/63 CAG repeats in ATXN3 to generate induced pluripotent stem cells (iPSCs), which will be a good model for studying SCA3.

Generation of two induced pluripotent stem cell lines, GZHMCi009-A and GZHMCi010-A, derived from peripheral blood mononuclear cells of two SCA3 patients with 14/74 CAG repeats of the ATXN3 mutation.

Spinal cerebellar ataxia type 3 (SCA3), also known as Machado-Joseph disease, is the result of abnormal repeat amplification of CAG of the ATXN3 gene. It is one of the main types of autosomal dominant ataxia, with motor symptoms of cerebellar ataxia, mainly accompanied by non-motor symptoms, such as ocular symptoms, psychiatric symptoms, and nutritional disorders. Currently, no effective treatment is available for patients with SCA3. The construction of induced pluripotent stem cells (iPSCs) from two SCA3 patients (14/74 CAG repeats) will be an excellent tool for studying SCA3 disease mechanisms and for drug screening.

Epilepsy Combined With Multiple Gene Heterozygous Mutation.

The fast pace of gene discovery has resulted in groundbreaking advances in the field of epilepsy genetics. Clinical testing using comprehensive gene panels, exomes, or genomes is now increasingly available and has significantly increased the diagnostic yield for early-onset epilepsies and enabled precision medicine approaches. In this paper, we report a case of epilepsy in a pedigree. The proband had heterozygous mutations in KCNC1 (NM_001112741.1:c.959G>A, p. Arg320His), CAPN3 (NM_000070.2:c.526G>A, p. Val176Met), and NEFH (NM_021076.3:c. 2595 delC, p. Lys866Argfs*51). Sanger sequencing verification was consistent with the results of whole-exome sequencing. The KCNC1 mutation was a de novo mutation, and the CAPN3 and NEFH mutations were inherited from their father and mother, respectively. Based on the American College of Medical Genetics and Genomics (ACMG) guidelines, a heterozygous mutation was found for APOB (NM_000384.2: c.10579C > T, p. Arg3527Trp). The heterozygous mutation at this site was inherent in the pedigree. Coexpression analysis indicated that heterozygous mutations of KCNC1, CAPN3, NEFH, and APOB were closely related to the clinical phenotypes of the patient, and the clinical phenotypic heterogeneity of the disease may be the result of the interaction of multiple genes.

Chlamydia trachomatis infection in the genital tract is associated with inflammation and hypospermia in the infertile male of China / 亚洲男科学杂志(英文版)

Chlamydia trachomatis (CT) infection is the most prevalent sexually transmitted bacterial disease worldwide. However, unlike that in female infertility, the role of CT infection in male infertility remains controversial. The objective of this retrospective study was to explore the impacts of CT infection in the genital tract on sperm quality, sperm acrosin activity, antisperm antibody levels, and inflammation in a large cohort of infertile males in China. A total of 7154 semen samples were collected from infertile male subjects, 416 of whom were CT positive (CT+ group) and 6738 of whom were CT negative (CT- group), in our hospital between January 2016 and December 2018. Routine semen parameters (semen volume, pH, sperm concentration, viability, motility, morphology, etc.), granulocyte elastase levels, antisperm antibody levels, and sperm acrosin activity were compared between the CT+ and CT- groups. Our results showed that CT infection was significantly correlated with an abnormally low semen volume, as well as an increased white blood cell count and granulocyte elastase level (all P < 0.05) in the semen of infertile males; other routine semen parameters were not negatively impacted. The antisperm antibody level and sperm acrosin activity were not affected by CT infection. These findings suggested that CT infection might contribute to inflammation and hypospermia but does not impair sperm viability, motility morphology, and acrosin activity or generate antisperm antibodies in the infertile males of China.

Detection rate and clinical significance of regions of homozygosity in prenatal genetic diagnosis / 中华妇产科杂志

Objective:To detect the incidence and analyze the clinical significance of regions of homozygosity (ROH) through the single nucleotide polymorphism array (SNP array).Methods:The SNP array detection results of 5 116 pregnant women in the Third Affiliated Hospital of Guangzhou Medical University from January 2016 to December 2020 were retrospectively analyzed. The pregnant women with ROH (5 Mb as the threshold) were followed up to analyze the relationship between ROH and abnormal fetal phenotype. Whole exon sequencing was performed in 4 cases of consanguineous marriage to detect potential recessive causative genes in the ROH region.Results:(1) A total of 39 cases of ROH were detected, with a positive rate of 0.76% (39/5 116). Among them, 25 cases (64%, 25/39) were detected only on single chromosome, and chromosome 11 had the highest detection rate, suggesting the risk of uniparental disomy; fourteen cases (36%,14/39) were detected on multiple chromosomes, most commonly on chromosomes 11, 1, 3, 4 and 8. (2) The number of cases and detection rate of ROH detected by different prenatal diagnosis indicators were as follows: 12 cases (1.78%, 12/676) in pregnant women with abnormal non-invasive prenatal testing result, 12 cases (0.37%, 12/3 284) in pregnant women with ultrasound abnormality, 4 cases (4/4) in pregnant women with consanguineous marriage, 3 cases (0.92%, 3/326) in pregnant women with previous adverse pregnancy, 2 cases (1.15%, 2/174) in pregnant women with high risk of serology in screening, 2 cases (4.00%, 2/50) in pregnant women with abnormal fetal chromosomal karyotype, 2 cases (0.79%, 2/253) in pregnant women with advanced maternal age, 1 case (0.56%, 1/178) in pregnant women with related parental genetic factors and 1 case (0.58%, 1/171) in pregnant women with the other factors. (3) The follow-up results of 39 cases of prenatal ROH showed that there were 16 cases of term birth, 15 cases of termination of pregnancy, 2 cases of preterm births, 1 case of fetal death and 5 cases lost to follow-up.Conclusions:Chromosomal ROH phenomenon is not rare. By analyzing the detection rate of ROH in prenatal diagnosis, combined with the results of fetal phenotype and postpartum follow-up, the clinical characteristics of ROH are discussed, so as to better understand the relationship between ROH and its phenotype.

Effect of Tongxinluo capsule on improvement of cardiac function and expression of myocardial enzyme spectrum in patients with coronary heart disease after percutaneous coronary intervention / 中国医师进修杂志

Objective:To investigate the effects of Tongxinluo capsule on the improvement of cardiac function and the expression of myocardial enzyme spectrum in patients with coronary heart disease after percutaneous coronary intervention (PCI).Methods:One hundred patients with coronary heart disease after PCI who received treatment in Hebei Xianghe County People′s Hospitalfrom April 2018 to April 2020 were enrolled and randomly divided into the observation group and the control group. The control group was given conventional treatment, while the observation group was treated with Tongxinluo capsule on the basis of the control group. Patients in both groups were treated for 3 months. The normal clinical remission after treatment was observed in the two groups. The improvement of cardiac function index left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), left ventricular ejection fraction (LVEF) and myocardial enzyme index aspartate aminotransferase (AST), creatine kinase (CK), creatine kinase isoenzyme - MB (CK - MB), lactate dehydrogenase (LDH), troponin T (TnT) were compared between the two groups before and after the treatment.Results:After treated for 3 months, the total effective rate in the observation group was higher than that in the control group: 92.0%(46/50) vs. 76.0%(38/50), the difference was statistically significant ( χ2 = 4.76, P<0.05). After treated for 3 months, the levels of LVEDV and LVESV in two groups were decreased, and the level of LVEF in two groups was increased, and the levels of LVEDV and LVESV in the observation group were lower than those in the control group: (153.39 ± 8.35) ml/m 2 vs. (155.57 ± 9.32) ml/m 2, (103.49 ± 9.25) ml/m 2 vs. (109.65 ± 10.46) ml/m 2; the levels of LVEF in the observation group was higher than that in the control group: (58.14 ± 7.41)% vs. (54.59 ± 6.92)%, the differences were statistically significant ( P<0.05). After treated for 3 months, the levels of AST, CK, CK-MB, LDH, TnT in two groups were decreased, and the levels of above index in the observation group were lower than those in the control group: (38.14 ± 7.28) U/L vs. (45.04 ± 8.12) U/L, (637.15 ± 75.25) U/L vs. (756.24 ± 85.24) U/L, (553.28 ± 53.14) U/L vs. (632.17 ± 62.81) U/L, (162.43 ± 15.41) U/L vs. (181.74 ± 19.25) U/L, (0.32 ± 0.15) μg/L vs. (0.39 ± 0.11) μg/L, the differences were statistically significant ( P<0.05). Conclusions:The application of Tongxinluo capsule in patients with coronary heart disease after PCI can effectively alleviate clinical symptoms, improve cardiac function, and regulate the enzyme activity of the body.

Study on single-incision laparoscopic appendectomy with conventional laparoscopic instruments / 中国医师进修杂志

Objective:To study the feasibility for transumbilical single-incision laparoscopic appendectomy (SILA) with conventional laparoscopic instruments, and compare SILA with the traditional three-port laparoscopic appendicectomy (LA).Methods:The clinical data of 113 patients with acute appendicitis from January 2018 to August 2020 in the Second Hospital of Jiaxing City were retrospectively analyzed. Among them, 61 patients received traditional three-port LA (three-port LA group), 52 patients received transumbilical SILA (SILA group). The operative time, intra-operative blood loss, surgical complications, length of hospital stay and hospitalization costs were recorded. Pain score 6 h after operation was assessed by visual analogue scale. C-reactive protein (CRP) on the first day after operation was detected. All patients were followed up for 1 month.Results:In 2 groups, all patients did not suffered from the conversion to open operation or multi-port method, massive bleeding and accessory injury during the operation. Moreover, severe pain, severe incision infection, residual abdominal abscess and incisional hernia did not occur. Patients in SILA group had more hidden abdominal scars. there were no statistical differences in operative time, intra-operative blood loss and CRP on the first day after operation between 2 groups ( P>0.05); the rate of moderate pain degree 6 h after operation, length of hospital stay and hospitalization costs in SILA group were significantly lower than those in three-port LA group: 15.38% (8/52) vs. 34.43% (21/61), (3.63 ± 1.22) d vs. (4.31 ± 1.38) d and (8 802 ± 1 466) yuan vs. (9 559 ± 1 617) yuan, and there were statistical differences ( P<0.05 or <0.01). Conclusions:The transumbilical SILA with conventional laparoscopic instruments is safe and feasible, the scar is more difficult to see, the cosmetic effect is much clearer, and the length of hospital stay is shorter.

Study on the relationship between serum Omentin-1 and metabolic parameters in patients with abdominal obesity / 中国综合临床

Objective:To investigate the level of serum Omentin-1 in subjects with abdominal obesity, and to analyze the influencing factors of Omentin-1 and its relationship with body fat distribution, insulin resistance and metabolic parameters.Methods:A retrospective case-control study was conducted to analyze the clinical data of one hundred and fifty adults with abdominal obesity (BMI≥28 kg/m 2) who were randomly selected from Obesity Multidisciplinary Diagnosis and Treatment Center, Subei People's Hospital from Januray 2018 to December 2021. Meanwhile, 150 healthy adults were enrolled as a normal control group. Fasting serum Omentin-1 and glucose metabolism were measured, and homeostasis model assessment of insulin resistance index (HOMA-IR) was calculated. Body fat composition was measured by bioelectrical impedance analysis. The relationship between Omentin-1 and other variables were presented by the Pearson correlation coefficients. Stepwise regression model was used to analyze the influencing factors of Omentin-1. Results:The serum omentin-1 level of patients with abdominal obesity was (36.97±6.99) μg/L, that of normal control group was (72.35±6.09) μg/L. The difference between the two groups was statistically significant ( t=46.69, P<0.001). The body fat level of patients with abdominal obesity was (43.40±14.59) kg, that of normal control group was (13.78±4.13) kg. The difference between the two groups was statistically significant ( t=23.93, P<0.001). The fasting insulin of patients with abdominal obesity was 29.05 (22.01,34.60) pmol/L, that of normal control group was 127.90 (84.08,201.45) pmol/L. The difference between the two groups was statistically significant( Z=14.75, P<0.001). The HOMA-IR of patients with abdominal obesity was 0.87 (0.68,1.05), that of normal control group was 4.19 (2.77,7.31). The difference between the two groups was statistically significant ( Z=14.75, P<0.001). Pearson linear correlation analysis showed that serum omentin-1 levels were negatively correlated with BMI, waist circumference, waist-hip ratio (WHR), body fat, visceral Fat area (VFA), HOMA-IR, triglyceride, total cholesterol and low density lipoprotein cholesterol ( r=-0.825, -0.843, -0.756, -0.777, -0.835, -0.583, -0.429, -0.353, -0.503, -0.938, all P<0.001). Whereas, a significantly positive correlation was found between serum Omentin-1 levels and high density lipoprotein cholesterol ( r=0.528, P<0.001). Omentin-1 concentrations were not related to age or gender ( r=-0.093, -0.040; P=0.669, 0.489). In multiple linear stepwise regression analysis, only VFA remained significantly associated with Omentin-1 ( β=-0.026, t=-2.250, P=0.026). Conclusion:The level of serum omentin-1 in patients with abdominal obesity was significantly lower than that in normal subjects, and it was closely related to body fat distribution and insulin resistance. VFA is an independent influencing factor of serum omentin-1, which can be used as a biomarker of abdominal obesity related metabolic disorders.

Generation of induced pluripotent stem cell GZHMCi005-A from amniotic fluid-derived cells with duplication of chromosome 8p.

Cases in which the duplication of chromosome 8p (dup 8p) is observed are characterized by facial dysmorphism, agenesis/hypoplasia of the corpus callosum, heart defects and severe mental retardation. The frequency of dup 8p cases is higher compared to other chromosomes because of the Non-allelic homologous recombination (NAHR) between two segmental duplication regions (SDs) containing olfactory receptor gene clusters, REPD (repeat-distal) and REPP (repeat-proximal), located in chromosome 8p23.1. Here we generated a human iPSC line from a patient's amniotic fluid cells with a 18 Mb duplication in 8p23.3p22, which will serve as useful tools for studying dup 8p syndrome.

Association of muscle mass loss with atherosclerosis in elderly patients with type 2 diabetes / 中华内分泌代谢杂志

Objective:To investigate the association of muscle mass loss with atherosclerosis in elderly patients with type 2 diabetes mellitus(T2DM).Methods:A total of 322 patients with T2DM aged≥60 years old were divided into muscle mass loss group( n=152) and non-muscle mass loss group( n=170) according to their appendicular skeletal muscle mass index(ASMI). All participants underwent physical examination, dual-energy X-ray absorptiometry check, carotid and lower extremity ultrasound, as well as laboratory tests. Results:Among 322 patients, 49(15.22%) patients were suffered from sarcopenia and 152(47.2%) patients with reduced muscle mass. The carotid and lower extremity atherosclerosis grades in the muscle mass loss group were significantly higher than those in the non-muscle mass loss group( P<0.05), with lower body mass index(BMI), T-score, ASMI, uric acid, and homeostasis model assessment of insulin resistance index( P<0.05). Multivariate logistic regression analysis showed that carotid atherosclerosis and lower extremity atherosclerosis were risk factors for muscle mass loss while BMI and 25-(OH)D 3 were protective factors for muscle mass loss. There existed a consistency in carotid atherosclerosis grade and lower extremity atherosclerosis grade of elderly patients with T2DM( P<0.01). Conclusion:Atherosclerosis has a predictive value for early sarcopenia in elderly patients with T2DM.

Research progress of iron metabolism in phenotype modification of β-thalassemia / 中华医学遗传学杂志

β-thalassemia is a type of inherited hemolytic anemia caused by decreased globin production due to defect of the HBB gene. The pathogenesis of the disease is imbalance of α/β globin chains. The excess of α-globin chains will form hemichromes which can damage red blood cell membranes and lead to hemolysis, ineffective erythropoiesis, and secondary iron overload. Iron overload in turn can cause complications such as growth retardation, liver cirrhosis, cardiac insufficiency, and aggravate the disease phenotype. In recent decades, genes participating in iron metabolism have been discovered, and the mechanism of iron metabolism in the development of thalassemia has gradually been elucidated. Subsequently, by manipulating the expression of key genes in iron metabolism such as hepcidin and transferrin receptor, researchers have revealed that iron restriction can improve ineffective hematopoiesis and iron overload, which may provide a potential approach for the treatment of thalassemia. This article reviews the progress of research on iron metabolism-related genes and related pathways in β-thalassemia.

Clinical practice guidelines for the diagnosis of regions of homozygosity and uniparental disomy / 中华医学遗传学杂志

The overall prevalence of uniparental disomy (UPD) across all chromosomes was estimated to be around one birth in 2000. To date, more than 4170 UPD cases have been registered. UPD for chromosomes 6, 7, 11, 14, 15, and 20 can result in clinically recognizable imprinting disorders due to abnormal levels of imprinted gene expression. For other chromosomes, the clinical consequences associated with UPD are not apparent, unless when a recessive genetic disorder is unmasked by UPD or regions of homozygosity (ROH). A clinical practice guideline will assist in strengthening the precise analysis and interpretation of the clinical significance of ROH/UPD. This guideline summarizes the conception, mechanism and clinical consequences of ROH/UPD, as well as the principles for data analysis, with an aim to standardize the clinical application and data interpretation.

Clinical practice guidelines for the diagnosis of regions of homozygosity and uniparental disomy / 中华医学遗传学杂志

The overall prevalence of uniparental disomy (UPD) across all chromosomes was estimated to be around one birth in 2000. To date, more than 4170 UPD cases have been registered. UPD for chromosomes 6, 7, 11, 14, 15, and 20 can result in clinically recognizable imprinting disorders due to abnormal levels of imprinted gene expression. For other chromosomes, the clinical consequences associated with UPD are not apparent, unless when a recessive genetic disorder is unmasked by UPD or regions of homozygosity (ROH). A clinical practice guideline will assist in strengthening the precise analysis and interpretation of the clinical significance of ROH/UPD. This guideline summarizes the conception, mechanism and clinical consequences of ROH/UPD, as well as the principles for data analysis, with an aim to standardize the clinical application and data interpretation.

Generation of induced pluripotent stem cell GZHMCi002-A from peripheral blood mononuclear cells with APOB mutation.

Apolipoprotein (apo) B is a large, amphipathic glycoprotein which plays an important role in human lipoprotein metabolism. The 43-kb APOB gene located on the short arm of human chromosome 2 and consisted of 29 exons, mutations in the APOB gene can give rise to either hypo- or hypercholesterolemia. We used peripheral blood mononuclear cells (PBMCs) from a volunteer carrying the APOB mutation (c.10579C>T, p.Arg3527Trp) located in exon 9 to establish induced pluripotent stem cells (iPSC), which will be an effective means to reveal the key biologically relevant metabolic mechanisms, a powerful tool for medicine selection and related research.

Generation of induced pluripotent stem cell GZHMCi001-A and GZHMCi001-B derived from peripheral blood mononuclear cells of epileptic patients with KCNC1 mutation.

Myoclonus Epilepsy and Ataxia due to Potassium channel mutation (MEAK) is a rare epilepsy caused by changes in the structure and function of potassium channels due to mutations in the potassium voltage-gated channel subfamily C member 1 (KCNC1) gene. MEAK is one of the progressive myoclonus epilepsy (PME), and there are few studies on MEAK pathogenesis and targeted drugs. Here, we used peripheral blood from MEAK patients with KCNC1 (c.959G > A) gene mutation to establish induced pluripotent stem cells (iPSC). The iPSC of KCNC1 mutation established by us is a powerful tool for related research.

Multivariate analysis of muscle mass loss in patients with type 2 diabetes / 中华内分泌代谢杂志

Objective:To explore the related factors of muscle mass loss in patients with type 2 diabetes mellitus, and to provide evidence for prevention of sarcopenia in type 2 diabetic patients.Methods:A cross-sectional survey was used to select type 2 diabetic patients admitted to the Department of Endocrinology, Affiliated Hospital of Qingdao University from January 2019 to August 2019. All subjects underwent dual-energy X-ray absorptiometry check. According to the diagnostic criteria of the Asian Working Group for Sarcopenia (AWGS), the subjects were divided into a muscle reduction group and a non-muscle reduction group. Data including age, gender, body mass index, course of disease, blood index, urinary albumin-creatinin ration (UACR), and appendicular skeletal muscle index (ASMI) were collected. The relevant clinical data of viscera fat/subcutaneous fat (VAT/SAT), percentage of abdominal fat/percentage of hip fat (A/G), grip strength, and pace were analyzed for the related factors of muscle mass loss in type 2 diabetic patients.Results:A total of 369 patients with type 2 diabetes were enrolled, including 42 patients with sarcopenia (an incidence rate of 11.38%), and 155 patients with reduced muscle mass (an incidence of 42.01%). Age, systolic blood pressure, high density lipoprotein-cholesterol, UACR, VAT/SAT, and A/G in the muscle reduction group were higher than those in the non-muscle reduction group ( P<0.05). The parameter of body mass index, homeostasis model assessment for insulin resistance, estimated glomerular filtration rate (eGFR), ASMI, and grip strength were lower in the muscle reduction group than in the non-muscle reduction group ( P<0.05). Multivariate logistic regression analysis showed age, UACR, A/G, and VAT/SAT were risk factors for muscle loss, body mass index, eGFR, and grip strength were protective factors for muscle mass reduction. Conclusion:It is of great practical significance for type 2 diabetic patients, especially those with advanced age, central obesity, low body mass index, low grip strength, low glomerular filtration rate, and high UACR to perform an early screening and to begin an early intervention.

Contrast-enhanced ultrasound combined with mathematical pattern identify intrahepatic cholangiocarcinoma and prophase of bacterial hepatic abscess / 中华超声影像学杂志

Objective:To explore the application of contrast-enhanced ultrasound (CEUS) combined with mathematical model in differential diagnosis of intrahepatic cholangiocarcinoma (ICC) and prophase of bacterial hepatic abscess (PBHA).Methods:Fifty-one ICC patients (ICC group)and 46 PBHA patients(PBHA group) from January 2010 to April 2019 in Henan Provincial People′s Hospital were analyzed retrospectively. All ICC patients and 41 PBHA patients were confirmed by surgery or puncture pathology, and 5 PBHA patients were confirmed by clinical follow-up. The characteristic information of ultrasound images of the lesions were analyzed.Partial least squares-discriminant analysis (PLS-DA) was used to establish ICC and PBHA mathematical models, and Monte Carlo simulation was used to verify their accuracy. Based on PLS-DA modeling method, variable importance in the projection (VIP) was used to obtain the variables that had a strong influence on identifying the two from all variables.Results:There were significant differences between ICC group and PBHA group in lesion sites, complications, internal characteristics and clearance characteristics of contrast-enhanced arterial phase (all P<0.05). The average positive and negative predictive values of PLS-DA were 92.30% and 91.17%, and the average positive and negative predictive values in the prediction set were 100% and 94.11%, respectively.Variables with VIP value >1 included X2 (complication), X4, X5, X7, X8, X9 (CEUS enhanced features), X10, X11 (clearing features), and these variables could be used as important indicators for differential diagnosis between ICC and PBHA. Conclusions:PLS-DA method based on CEUS parameters can construct a differential diagnosis model for ICC and PHBA, which is expected to provide a valuable and robust diagnostic method for these two diseases which are easily confused and lack of specific imaging manifestations.

Generation of four induced pluripotent stem cell lines, GZWWTi001-A, GZWTZi001-A, GZWXYi001-A, and GZWXDi001-A, derived from peripheral blood mononuclear cells from a family with asparagine synthetase deficiency.

Asparagine synthetase (ASNS) deficiency (ASNSD; MIM #615574) is a rare neurodevelopmental disorder caused by mutations in the ASNS gene. The ASNS gene maps to cytogenetic band 7q21.3 and is 35 kb long. ASNSD is characterised by congenital microcephaly, severely delayed psychomotor development, seizures, and hyperekplexic activity. Here, we reported a family with compound heterozygous mutations in ASNS (NM_001178076:c.551C>T; c. 944A>C) and established induced pluripotent stem cells (iPSCs) from blood samples. To date, limited functional data have been reported to explain the underlying pathophysiology of ASNSD; therefore, iPSCs from these patients may be powerful tools for studying disease mechanisms.

CRISPR/Cas9 gene correction of HbH-CS thalassemia-induced pluripotent stem cells.

Haemoglobin (Hb) H-constant spring (CS) alpha thalassaemia (- -/-αCS) is the most common type of nondeletional Hb H disease in southern China. The CRISPR/Cas9-based gene correction of patient-specific induced pluripotent stem cells (iPSCs) and cell transplantation now represent a therapeutic solution for this genetic disease. We designed primers for the target sites using CRISPR/Cas9 to specifically edit the HBA2 gene with an Hb-CS mutation. After applying a correction-specific PCR assay to purify the corrected clones followed by sequencing to confirm the mutation correction, we verified that the purified clones retained full pluripotency and exhibited a normal karyotype. This strategy may be promising in the future, although it is far from representing a solution for the treatment of HbH-CS thalassemia now.

Effect of liraglutide on glucagon secretion in obese type 2 diabetic patients / 中华内科杂志

Objective To investigate the effect of liraglutide on glucagon release in obese type 2 diabetes (T2DM). Methods A multi-center, prospective, and self-comparison study was conducted in four hospitals in Qingdao. Twenty-four patients with T2DM were selected and treated with liraglutide for 12 weeks. Glucagon levels before and after treatment were detected before and 30 min, 60 min and 120 min after meals. Results After 12 weeks of treatment, the overall level of glucagon decreased, in which the differences in glucagon levels at 30 min [(220±79) ng/L vs. (203±77) ng/L, P<0.05] and 60 min [(248±119) ng/L vs. (203±82)ng/L, P<0.05] reached significance, respectively, comparing to those before treatment. The area under the curve of glucagon after treatment was significantly lower than that before treatment (438±190 vs. 389 ± 153, P<0.05). In contrast, after treatment, the overall level of C-peptide increased, especially the levels at 30 min [(1.53±1.02) nmol/L vs.(2.03±1.29) nmol/L ], 60 min [(1.93±1.19) nmol/L vs. (2.48±1.75) nmol/L] and 120 min [(2.36±1.47) nmol/L vs. (2.96±1.84) nmol/L], all P<0.05. The area under C-peptide curve increased significantly (3.6±2.2 vs. 4.6±2.9, P<0.05). Fasting plasma glucose, postprandial 2 h plasma glucose and glycosylated hemoglobin A1c were all lower than before, and the differences were statistically significant (P<0.05). Waist circumference and body mass index were significantly lower than before (P<0.05). The amount of insulin used for the treatment decreased by approximately 55.1% compared with that before liraglutide, and the difference was statistically significant (P<0.05). Conclusions Liraglutide inhibits glucagon secretion and lowers blood glucose. It can also reduce body weight, improve islet cell function and reduce insulin use in T2DM.