Association between single nucleotide polymorphisms, TGF-ß1 promoter methylation, and polycystic ovary syndrome.

BACKGROUND: Polycystic ovarian syndrome (PCOS) is a common endocrine and metabolic disease in women. Hyperandrogenaemia (HA) and insulin resistance (IR) are the basic pathophysiological characteristics of PCOS. The aetiology of PCOS has not been fully identified and is generally believed to be related to the combined effects of genetic, metabolic, internal, and external factors. Current studies have not screened for PCOS susceptibility genes in a large population. Here, we aimed to study the effect of TGF-ß1 methylation on the clinical PCOS phenotype. METHODS: In this study, three generations of family members with PCOS with IR as the main characteristic were selected as research subjects. Through whole exome sequencing and bioinformatic analysis, TGF-ß1 was screened as the PCOS susceptibility gene in this family. The epigenetic DNA methylation level of TGF-ß1 in peripheral blood was detected by heavy sulfite sequencing in patients with PCOS clinically characterised by IR, and the correlation between the DNA methylation level of the TGF-ß1 gene and IR was analysed. We explored whether the degree of methylation of this gene affects IR and whether it participates in the occurrence and development of PCOS. RESULTS: The results of this study suggest that the hypomethylation of the CpG4 and CpG7 sites in the TGF-ß1 gene promoter may be involved in the pathogenesis of PCOS IR by affecting the expression of the TGF-ß1 gene. CONCLUSIONS: This study provides new insights into the aetiology and pathogenesis of PCOS.

Morphological and radiological features of congenital muscular hypertrophy of the upper limb: experience from a tertiary institution.

Congenital muscular hypertrophy is a rare overgrowth disorder in the phosphatidylinositol-3-kinase related spectrum. In the past 3 years, ten patients with 11 limbs involved were treated in our centre. The aim of the study was to describe the clinical and radiological deformities of these patients. We documented the characteristic clinical morphological changes, such as hypertrophy, loss of wrist flexion, thumb hyperabduction, finger deviation and skin crease changes in the palm. Radiologically, the mean first metacarpal radial deviation angle of the affected side measured 55° (range 34 to 67) compared to the normal contralateral side 42° (range 32 to 53). The mean intermetacarpal space ratio was 1.2 (range 1.1 to 1.4) and the mean palm width ratio was 1.2 (range 1.1 to 1.3). In this study, we were able to further characterize the radiological and morphological changes of congenital muscular hypertrophy of upper limbs, which would be helpful for establishing the diagnosis and monitor treatment of this rare condition.Level of evidence: IV.

Triplication and tetraplication of the thumb: a modification of the Rotterdam classification.

We proposed a modification of the Rotterdam classification for thumb triplication and tetraplication. Twenty-one patients were included (24 cases of thumb triplication and four cases of tetraplication). These were analysed and classified according to a modification of the Rotterdam classification involving three steps; from the radial to ulnar side, we first identified each thumb on radiographs and gross appearance to divide into triplication or tetraplication. Second, we define the levels of duplication and established the nomenclature. Third, the aberrant features and their location were assigned for each thumb, again from radial to ulnar side. A surgical algorithm was also proposed. This modified classification may be helpful for characterizing the rare conditions of thumb triplication and tetraplication for use in patient and management and communication between surgeons.Level of evidence: III.

Assessment of the risk of unexplained recurrent spontaneous abortion based on the proportion and correlation of NK cells and T cells in peripheral blood.

BACKGROUND: Unexplained recurrent spontaneous abortion (URSA) is difficult to diagnose and treat clinically due to its unknown causeOBJECTIVE: Changes in natural killer (NK) cells, T lymphocytes, and Th1(IFNγ)/Th2(IL-4) cytokines were investigated in the peripheral blood of patients with URSA to examine the pathogenesis, clinical diagnosis, and inform potential treatment strategies for this condition. METHODS: For this study, we selected patients with URSA as the case group and included normal women in the control group. Flow cytometry was performed to detect lymphocytes and cytokines in the peripheral blood of all subjects. RESULTS: The proportion of NK cells, Th1 cells, and the Th1/Th2 ratio were significantly higher in the URSA group compared to the control group; whereas the proportion of CD3+T cells was lower. Pairwise correlation analysis revealed a positive correlation between the percentage of NK cells and CD3+T cells, as well as CD3+CD4+T cells. Canonical correlation analysis indicated a significant correlation between NK cells and T cells, including their subgroups. CONCLUSION: Patients with URSA have immune balance disorders, characterised by an increased proportion of peripheral blood NK cells, Th1, and Th1/Th2 ratio along with a decreased proportion of CD3+T cells. The proportion of NK cells and CD3+T may serve as predictive factors for URSA, while NK cells are closely related to the regulation of CD3+T cells and their subsets. By regulating the level of IFN-γ, NK cells can influence the proportion of CD3+T cells and induce a Th1 (IFNγ)/Th2 (IL-4) imbalance.

Comparison Study of 5-flap Z-Plasty and Double Z-Plasty for Interdigital Pocket Web Contractures.

PURPOSE: The aim of this study was to compare the improvement in abduction and change in web slope between 5-flap z-plasty and double z-plasty for post-burn or post-trauma interdigital pocket web contractures. METHODS: We retrospectively reviewed 16 webs in 11 patients with post-burn or post-trauma interdigital pocket web contractures. Seven patients underwent 5-flap z-plasty and 4 double z-plasty. The abduction and slope angles of the affected fingers were measured before surgery and every 3 months after surgery. RESULTS: The follow-up time ranged from 12 to 36 months. The postoperative abduction angle improved in both groups. There was an increase of 19° ± 4° in the mean abduction angle after surgery in the 5-flap z-plasty group compared with a mean of 12° ± 2° in the double z-plasty group; the postoperative decrease in the web slope angle was 30° ± 5° and 22° ± 2°, respectively. All patients were able to actively move the affected webs and fingers with no pain or other discomfort. CONCLUSIONS: Five-flap z-plasty can be considered a better alternative to double z-plasty for interdigital pocket web contractures based on the improvement in abduction angle and changes in the slope angle. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV.

Nonvascularized Iliac Crest Bone Graft for Reconstruction of the First Metacarpal in Type IIIB Thumb Hypoplasia: A Radiographic Follow-Up Study.

PURPOSE: To present a radiographic follow-up study of the use of a nonvascularized iliac crest bone graft as a treatment for modified Blauth type IIIB thumb hypoplasia. METHODS: From January 2015 to December 2019, nonvascularized iliac crest bone grafts were used to reconstruct the first metacarpal in 23 cases with type IIIB thumbs. The average follow-up duration was 1.9 years (range 1.0-3.9 years). We evaluated the patients' serial x-rays and measured the width and length changes of the reconstructed first metacarpals. RESULTS: Survival of the graft, judged radiographically, was achieved in 20 cases (20/23, 87%), with an average reduction of 33% in the width of the graft. Shortening of the graft (average 2.3 mm, range 0.1-5.6 mm) was noted in 11 of the 20 cases, and an increase in the length of the graft (average 4.2 mm, range 0.7-8.6 mm) was observed in the other 9. Bone graft failure (3/23, 13%) occurred in 3 cases because of significant bone resorption. CONCLUSIONS: For type IIIB thumb hypoplasia, a nonvascularized iliac crest bone graft was a feasible method to reconstruct the first metacarpal, with a 13% risk of graft failure, 33% average reduction in graft width, and 55% reduction in graft length. However, in cultures that value the preservation of a 5-digit hand, this technique may provide an alternative to excision with index pollicization. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV.

A missense variant in NCF1 is associated with susceptibility to unexplained recurrent spontaneous abortion.

Unexplained recurrent spontaneous abortion (URSA) is a major concern in reproductive medicine. Neutrophil cytosolic factor 1 (NCF1) polymorphisms leading to low production of reactive oxygen species (ROS) are strongly associated with autoimmune diseases. We investigated the association of the missense single nucleotide polymorphism (SNP) rs201802880 (NCF1-339) in NCF1 with URSA and explored its function. We performed NCF1-339 SNP genotyping of samples from 152 Chinese patients with URSA and 72 healthy controls using nested PCR and TaqMan assays. ROS production and RELA (NF-κB subunit) expression in the blood of participants with different NCF1-339 genotypes were determined. The frequencies of the wild-type (GG) and mutant (GA) genotypes remarkably differed between the URSA and control groups. The mutant genotype was associated with an increased risk of recurrent abortion. Furthermore, ROS levels in the URSA group with the GG genotype were significantly higher than those in the group with the GA genotype (p < 0.05). RELA expression in URSA patients with the GA genotype was considerably higher than that in control individuals with the GG genotype. These findings indicate that mutations in NCF1 may increase the risk of URSA via the NADP/ROS/NF-κB signaling pathway, which has implications for the diagnosis and treatment of URSA.

[Soft tissue balance technique by flexor pollicis longus tendon transfer for Wassel â £-D thumb duplication in children].

Objective: To investigate the effectiveness of soft tissue balance technique by flexor pollicis longus (FPL) tendon transfer for Wassel Ⅳ-D thumb duplication in children. Methods: A clinical data of 14 children with Wassel Ⅳ-D thumb duplication met the selection criteria between January 2017 and January 2021 was retrospectively analyzed. There were 5 boys and 9 girls with an average age of 21.6 months (range, 18-35 months). Ten cases were left hand deformity and 4 cases were right hand deformity. During operation, the radial thumb was excised, and the FPL tendon of the radial thumb was used to reconstruct the soft tissue balance of the ulnar thumb. Postoperative evaluation included the range of motion (ROM) of passive flexion and extension of the interphalangeal joint (IP) and metacarpophalangeal joint (MCP), the alignments of the IP and MCP, the percentage of the width of the nail plate and the circumference of the thumb at the level of the IP to contralateral thumb. Results: All operations were completed successfully, and all incisions healed by first intention. The children were followed up 12-36 months (mean, 21.7 months). At last follow-up, the ROM of passive flexion and the deviation of the IP, and the deviation of the MCP significantly improved when compared with those before operation (P<0.05); the ROM of passive extension of the IP and the ROM of passive flexion of the MCP did not significantly improve when compared with those before operation (P>0.05). The ROMs of passive extension of the MCP were 0° before and after operation. The width of the nail plate was 76.6%±4.1% of the unaffected side, and the circumference of the thumb at the level of the IP was 92.0%±9.1% of the unaffected side. Conclusion: The soft tissue balance technique by FPL tendon trasfer can effectively correct the alignment of the Wassel Ⅳ-D thumb duplication in children, and maintain the correction effect effectively, but further follow-up and comprehensive evaluation are needed to investigate the long-term effectiveness.

Identification of Novel FBN2 Variants in a Cohort of Congenital Contractural Arachnodactyly.

Congenital contractural arachnodactyly (CCA) is a rare autosomal dominant disorder of connective tissue characterized by crumpled ears, arachnodactyly, camptodactyly, large joint contracture, and kyphoscoliosis. The nature course of CCA has not been well-described. We aim to decipher the genetic and phenotypic spectrum of CCA. The cohort was enrolled in Beijing Jishuitan Hospital and Peking Union Medical College Hospital, Beijing, China, based on Deciphering disorders Involving Scoliosis and COmorbidities (DISCO) study (http://www.discostudy.org/). Exome sequencing was performed on patients' blood DNA. A recent published CCA scoring system was validated in our cohort. Seven novel variants and three previously reported FBN2 variants were identified through exome sequencing. Two variants outside of the neonatal region of FBN2 gene were found. The phenotypes were comparable between patients in our cohort and previous literature, with arachnodactyly, camptodactyly and large joints contractures found in almost all patients. All patients eligible for analysis were successfully classified into likely CCA based on the CCA scoring system. Furthermore, we found a double disease-causing heterozygous variant of FBN2 and ANKRD11 in a patient with blended phenotypes consisting of CCA and KBG syndrome. The identification of seven novel variants broadens the mutational and phenotypic spectrum of CCA and may provide implications for genetic counseling and clinical management.

Activating PIK3CA postzygotic mutations in segmental overgrowth of muscles with bone involvement in the body extremities.

Segmental overgrowth of the skeletal muscles with bone involvement in body extremities, predominantly affecting the upper limb, is an extremely rare condition with only 40-50 affected children described clinically. The molecular pathogenesis of this disorder remains largely unclear except for the identification of a somatic PIK3CA mutation in each of the six patients genetically tested, all restricted to upper limbs in the literature. This study aimed to further characterize the molecular defects for patients affected with segmental overgrowth of the skeletal muscles by analyzing a 9-gene panel selected from the PI3K/AKT/mTOR pathway and genes associated with other related conditions. Nineteen unrelated patients were chosen for this study, comprising ten upper limb (nine unilateral and one bilateral) and nine lower limb (eight unilateral and one bilateral) cases with variable bone involvement. In each case, an activating PIK3CA mutation (p.E110del, p.N345K, p.E542K, p.E545K, p.H1047R, or p.H1047L) was identified in the affected muscle tissue with variant allele frequencies ranging from 13.88 to 30.43%, while no mutation was detected in the paired peripheral blood sample, indicating somatic mosaicism. All detected mutations were limited to PIK3CA and were previously reported in other overgrowth syndromes currently categorized under the PIK3CA-Related Overgrowth Spectrum (PROS). Our study provides strong molecular evidence that isolated segmental overgrowth of the skeletal muscle with bone involvement is a subtype of PROS. Our findings expand the PROS clinical presentations with a newly molecularly classified condition and can provide guidance in clinical and molecular diagnosis and treatment for patients with this condition.

Single nucleotide polymorphisms in binding site of miRNA-135a and targeted gene IRS2 are correlated with multiple clinical features of PCOS: A study in Chinese women.

BACKGROUND: The etiology of polycystic ovary syndrome (PCOS) remains unclear with highly heterogeneous clinical manifestations, recently growing evidence revealing genetic variants play a crucial part in its pathogenesis. OBJECTIVE: This study aimed to examine the correlation between SNPs in miRNA-135a's binding site of targeted gene IRS2 and clinical manifestations of PCOS in Chinese females. METHOD: A total of 126 Chinese women with PCOS and 109 healthy women were enrolled, divided into 4 groups based on different clinical features of hyperandrogenemia (HA), insulin resistance (IR), polycystic ovary morphology (PCOM) and obesity. We analyzed 2 single nucleotide polymorphisms (SNPs) of the IRS2 gene (rs2289046 and rs1865434) and clinical features' laboratory measurements such as sex hormone, fasting plasma glucose (FPG), fasting plasma insulin (FINS). RESULTS: Located in miRNA-135a binding site of IRS2 gene, the rs2289046's triple genotypes distribution showed a significant difference between PCOS/control group and PCOM/non-PCOM group (P< 0.05) while the rs1865434's triple genotype distribution showed a significant difference between obesity/non-obesity group (P< 0.05). CONCLUSION: The results revealed the two SNPs as rs2289046 and rs1865434 in the IRS-2 binding region of miRNA-135a have correlations with the clinical features of PCOS in Chinese population.

Deciphering the mutational signature of congenital limb malformations.

Congenital limb malformations (CLMs) affect 1 in 500 live births. However, the value of exome sequencing (ES) for CLM is lacking. The purpose of this study was to decipher the mutational signature of CLM on an exome level. We enrolled a cohort of 66 unrelated probands (including 47 families) with CLM requiring surgical correction. ES was performed for all patients and available parental samples. A definite molecular diagnosis was achieved in 21 out of 66 (32%) patients. We identified 19 pathogenic or likely pathogenic single-nucleotide variants and three copy number variants, of which 11 variants were novel. We identified four variants of uncertain significance. Additionally, we identified RPL9 and UBA2 as novel candidate genes for CLM. By comparing the detailed phenotypic features, we expand the phenotypic spectrum of diastrophic dysplasia and chromosome 6q terminal deletion syndrome. We also found that the diagnostic rate was significantly higher in patients with a family history of CLM (p = 0.012) or more than one limb affected (p = 0.034). Our study expands our understanding of the mutational and phenotypic spectrum of CLM and provides novel insights into the genetic basis of these syndromes.

Phenotypic and genetic spectrum of isolated macrodactyly: somatic mosaicism of PIK3CA and AKT1 oncogenic variants.

BACKGROUND: Isolated macrodactyly is a severe congenital hand anomaly with functional and physiological impact. Known causative genes include PIK3CA, AKT1 and PTEN. The aim of this study is to gain insights into the genetics basis of isolated macrodactyly. RESULTS: We enrolled 24 patients with isolated macrodactyly. Four of them were diagnosed with Proteus syndrome based on skin presentations characteristic to this disease. Targeted next-generation sequencing was performed using patients' blood and affected tissues. Overall, 20 patients carry mosaic PIK3CA pathogenic variants, i.e. p.His1047Arg (N = 7), p.Glu542Lys (N = 6), p.Glu545Lys (N = 2), p.His1047Leu (N = 2), p.Glu453Lys (N = 1), p.Gln546Lys (N = 1) and p.His1047Tyr (N = 1). Four patients who met the diagnostic criteria of Proteus syndrome carry mosaic AKT1 p.Glu17Lys variant. Variant allele frequencies of these mosaic variants obtained through next-generation sequencing range from 10 to 33%. In genotype-phenotype correlation analysis of patients with PIK3CA variant, we found that patients with the macrodactyly of the lower limbs tend to carry PIK3CA variants located in the helical domain (P = 0.005). CONCLUSIONS: Mosaic PIK3CA and AKT1 variants can be found in all of our samples with isolated macrodactyly. Insights into phenotypic and genetic spectrum of isolated macrodactyly may be helpful in perusing a more precise and effective management of isolated macrodactyly.

Two-Stage "On-Top Plasty" for Thumb Polydactyly With a Floating Ulnar Thumb: 3 Case Reports.

We present 3 cases of thumb polydactyly treated with a 2-stage "on-top plasty" technique. For all the 3 cases, the ulnar thumbs floated in the first web with a skin pedicle, and the radial thumbs presented hypoplasia of different degree. We designed a 2-stage "on-top plasty" technique. The first operation included transplantation of the ulnar floating thumb and construction of skin tube containing the neurovascular bundle. When followed up after the first operation, there was no vascular compromise, and all union was achieved. At the time of the second operation, the skin tube was opened up, the neurovascular bundle was transposed to the reconstructed thumb, and the first web plasty was performed. At the final follow-up, 3 cases showed acceptable results both cosmetically and functionally.

A Cadaveric Study on the Accuracy of an Individualized Guiding Template to Assist Scaphoid Fixation Using Computed Tomography and 3-Dimensional Printing.

PURPOSE: The aim of this study was to evaluate the feasibility and accuracy of scaphoid screw guidewire placement using a computer-assisted-designed, and 3-dimensional-printed, surgical guiding template in cadaver wrists. METHODS: Computed tomography (CT) scans of 12 fresh-frozen cadaver wrists were performed and the data imported into a surgical planning system. A 3-dimensional skin surface template block with a guiding hole was generated from the CT data to allow a screw guidewire to be placed in the central third of the scaphoid. This 3-dimensional model was printed and then put back onto the wrist. A screw guidewire was inserted through the palmar guide hole into the intact scaphoid and then a postprocedure CT scan was obtained. These postprocedure data were introduced into the surgical planning system. Angular and linear deviation between the preprocedural simulation and the image of the guidewire was measured in the system to assess accuracy. RESULTS: Mean angular deviation was 3.85° ± 1.32° (range, 1.56°-5.35°) and linear deviations of the 12 specimens were less than 1.1 mm. No specimen required a repeat drilling to the scaphoid. All the screw guidewires were considered to be centrally placed in the scaphoid based on our criterion of central placement of the scaphoid screw. CONCLUSIONS: The use of a computer-assisted 3-dimensional-printed surgical guide template to assist screw guidewire placement into an intact scaphoid, mimicking a nondisplaced scaphoid fracture, showed acceptable accuracy in cadaver wrists. CLINICAL RELEVANCE: Our technique may provide a simple and effective method for the guidance of screw guidewire insertion in a nondisplaced scaphoid fracture surgery.