OBJECTIVES: The purinergic receptor P2X2 (P2RX2) encodes an ATP-gated ion channel permeable to Na⁺, K⁺, and especially Ca²⁺. Loss-of-function mutations in P2RX2 are known to cause autosomal dominant nonsyndromic deafness 41 (DFNA41), which manifests as high-frequency hearing loss, accelerated presbycusis, and increased susceptibility to noise-induced damage. Zebrafish, owing to their small size, rapid development, high fecundity, transparent embryos, and high gene conservation with humans, provide an ideal model for studying human diseases and developmental mechanisms. This study aims to generate a p2rx2 knockout zebrafish model using CRISPR/Cas9 gene editing system to investigate the effect of p2rx2 deficiency on the auditory system, providing a basis for understanding P2RX2-related hearing loss and developing gene therapy strategies. METHODS: Two CRISPR targets (sgRNA1 and sgRNA2) spaced 47 bp apart were designed within the zebrafish p2rx2 gene. Synthesized sgRNAs and Cas9 protein were microinjected into single-cell stage Tübingen (TU)-strain zebrafish embryos. PCR and gel electrophoresis verified editing efficiency at 36 hours post-fertilization (hpf). Surviving embryos were raised to adulthood (F0), tail-clipped, genotyped, and screened for positive mosaics. F1 heterozygotes were generated by outcrossing, and F2 homozygous mutants were obtained by intercrossing. Polymerase chain reaction (PCR) combined with sequencing verified mutation type and heritability. At 5 days post-fertilization (dpf), YO-PRO-1 staining was used to examine hair cell morphology and count in lateral line neuromasts and the otolith region. Auditory evoked potential (AEP) thresholds at 600, 800, 1 000, and 2 000 Hz were measured in nine 4-month-old wild type and mutant zebrafish per group. RESULTS: A stable p2rx2 knockout zebrafish line was successfully established. Sequencing revealed a 66 bp insertion at the first target site introducing a premature stop codon (TAA), leading to early termination of protein translation and loss of function. Embryos developed normally with no gross malformations. At 5 dpf, mutants exhibited significantly reduced hair cell density in the otolith region compared with wild type, although lateral line neuromasts were unaffected. AEP testing showed significantly elevated auditory thresholds at all 4 frequencies in homozygous mutants compared with wild type (all P<0.001), indicating reduced hearing sensitivity. CONCLUSIONS We successfully generated a p2rx2 loss-of-function zebrafish model using CRISPR/Cas9 technology. p2rx2 deficiency caused hair cell defects in the otolith region and increased auditory thresholds across frequencies, indicating its key role in maintaining zebrafish auditory hair cell function and hearing perception. The phenotype's restriction to the otolith region suggests tissue-specific roles of p2rx2 in sensory organs. This model provides a valuable tool for elucidating the molecular mechanisms of P2RX2-related hearing loss and for screening otoprotective drugs and developing gene therapies.
Animals ; Zebrafish/genetics* ; Receptors, Purinergic P2X2/deficiency* ; CRISPR-Cas Systems/genetics* ; Gene Knockout Techniques ; Phenotype ; Zebrafish Proteins/genetics* ; Disease Models, Animal

Objectives:To investigate the clinical efficacy of the distal tension-offloading cosmetic suture technique in reducing hypertrophic scar formation following open thyroidectomy.Methods:Clinical data and postoperative incision appearance of 138 patients undergoing open thyroidectomy at the Department of Thyroid and Neck Oncology of Tianjin Medical University Cancer Institute and Hospital, as well as the Department of Head and Neck Oncology of Tianjin Cancer Hospital Airport Hospital, from Aug 2023 to Jan 2024 was enrolled. Patients were devided into two groups based on the incision closure method: the distal tension-offloading cosmetic suture group (tension reduction group, 37 cases) and ordinary intradermal suture group (control group A, 55 cases evaluated 3 months post-surgery; control group B, 46 cases evaluated 6 months post-surgery). The Vancouver Scar Scale (VSS) was employed to assess the appearance of the wounds at 3 and 6 months post-surgery .Results:On 3 and 6 months post-surgery, the total VSS scores for patients in the tension reduction group were 2.8 ± 2.3 and 2.5 ± 2.5, respectively,while that in control group A on 3 months was 5.2 ± 3.0, and in group B on 6 months was 5.3 ± 3.4. The differences were statistically significant (all P<0.001). On 3 and 6 months post-surgery, the proportions of hypertrophic scars in the tension reduction group were 14% (5/37) and 11% (4/37), respectively , while in control group A it was 35% (19/55) , and in control group B was 35% (16/46) at 6 months, with differences being statistically significant ( P=0.024, 0.011 ). On 6 months post-surgery, 51 % (19/37) of patients in the tension reduction group achieved 'socially invisible aesthetic incisions', while only 15% (7/46) of patients in control group B achieved the same outcome ( P<0.01). Conclusion:Distal tension-offloading cosmetic suture significantly reduces the incidence of incision hypertrophic scars in open thyroid surgery.

Objectives:To investigate the clinical efficacy of the distal tension-offloading cosmetic suture technique in reducing hypertrophic scar formation following open thyroidectomy.Methods:Clinical data and postoperative incision appearance of 138 patients undergoing open thyroidectomy at the Department of Thyroid and Neck Oncology of Tianjin Medical University Cancer Institute and Hospital, as well as the Department of Head and Neck Oncology of Tianjin Cancer Hospital Airport Hospital, from Aug 2023 to Jan 2024 was enrolled. Patients were devided into two groups based on the incision closure method: the distal tension-offloading cosmetic suture group (tension reduction group, 37 cases) and ordinary intradermal suture group (control group A, 55 cases evaluated 3 months post-surgery; control group B, 46 cases evaluated 6 months post-surgery). The Vancouver Scar Scale (VSS) was employed to assess the appearance of the wounds at 3 and 6 months post-surgery .Results:On 3 and 6 months post-surgery, the total VSS scores for patients in the tension reduction group were 2.8 ± 2.3 and 2.5 ± 2.5, respectively,while that in control group A on 3 months was 5.2 ± 3.0, and in group B on 6 months was 5.3 ± 3.4. The differences were statistically significant (all P<0.001). On 3 and 6 months post-surgery, the proportions of hypertrophic scars in the tension reduction group were 14% (5/37) and 11% (4/37), respectively , while in control group A it was 35% (19/55) , and in control group B was 35% (16/46) at 6 months, with differences being statistically significant ( P=0.024, 0.011 ). On 6 months post-surgery, 51 % (19/37) of patients in the tension reduction group achieved 'socially invisible aesthetic incisions', while only 15% (7/46) of patients in control group B achieved the same outcome ( P<0.01). Conclusion:Distal tension-offloading cosmetic suture significantly reduces the incidence of incision hypertrophic scars in open thyroid surgery.

【Objective】 To improve the understanding and diagnosis and treatment level of ALK negative anaplastic large cell lymphoma (ALK-ALCL) by sharing the diagnosis and treatment process of a patient with ALK-ALCL treated in Hangzhou Bay Hospital of Ningbo. 【Methods】 The clinical data and diagnosis and treatment process of the patient were retrospectively analyzed, and relevant literature was reviewed. 【Results】 The patient was a young male, with recurrent gross hematuria and right low back pain as the initial symptoms.Imaging examination indicated bladder tumor.After resection, the tumor was reduced and confirmed to be ALK-ALCL.After chemotherapy and autologous hematopoietic stem cell transplantation, the patient’s condition continued to improve.During the follow-up, no recurrence was observed. 【Conclusion】 Primary ALK-ALCL in the bladder is very rare and prone to misdiagnosis and missed diagnosis in clinical practice.The successful diagnosis and treatment experience of this patient can provide clinical reference.

Objective:To investigate the clinical efficacy of labial mucosa graft coupled preputial island flap urethroplasty for recurrent penile curvature after hypospadias repair.Methods:The data of 35 patients with recurrent penile curvature after hypospadias repair who underwent labial mucosa graft coupled preputial island flap urethroplasty in four hospitals from January 2021 to March 2023 were retrospectively reviewed. The median age of the patients was 15(13, 18)years old, five of them had penile dehiscence, and four had urethrocutaneous fistula. All patients received labial mucosa graft coupled preputial island flap urethroplasty, the penile flexion was confirmed by artificial erection test, and the urethra is obliquely cut at the most obvious curvature of the penis. By removing the ventral fibrotic scar tissue, the length of the urethral defect was measured. The urethral plate was reconstructed with the same length of labial mucosa graft and an adjacent preputial island flap was transplanted to cover the reconstructed urethra plate to form a new urethral lumen. The inner urethral stent tube was extracted three days after the operation, and the outer urethral stent tube was removed three weeks after the surgery. Diagnostic urethral exploration was performed 5 weeks post-operation, uroflowmetry examination was performed seven weeks after the procedure, and the perioperative and postoperative follow-up data were analyzed.Results:All patients underwent a successful surgery, with the average duration of the procedure being (114.7±20.1)minutes. After mean follow-up of (15.5±5.9)months, no penile curvature occurred, and 32 patients were successfully repaired. Uroflowmetry examination was achieved in 21 patients at the seventh week after surgery showed their mean maximum flow rate was (18.4±3.7)ml/s. Three cases (8.6%, 3/35) developed urethrocutaneous fistula, who were repaired successfully 6 months after surgery.Conclusions:Utilizing the combination of labial mucosa grafts and preputial island flaps for urethroplasty has been proven to be a reliable and safe option for individuals with recurrent penile curvature following hypospadias repair, with a high success rate and few complications.

Objective To investigate the effect of the loss of myeloid-derived growth factor(Mydgf)on the transformation of cardiac fibroblasts into myofibroblasts after myocardial infarction(MI).Methods Two adult mouse groups,including a wild-type(WT)group and another group with Mydgf knockout(Mydgf-KO),were examined in the study.The mice in these two groups were tested for their cardiac function by measuring left ventricular ejection fraction(LVEF)and left ventricular fractional shortening(LVFS)(n=10).Quantitative real-time PCR(qRT-PCR)(n=3)was performed to determine the mRNA expression levels of myofibroblast markers,including α-smooth muscle actin(α-SMA),periostin(postn),type Ⅷ collagen(col8al),and connective tissue growth factor(ctgf).Western blot(n=3)was performed to verify the protein expression levels of α-SMA.MI modeling was performed on the WT and the Mydgf-KO mice.Postoperative LVEF and LVFS(n=10)were then measured.The hearts were harvested and Masson staining was performed to determine the infarcted area(n=10).The heart samples of Mydgf-KO and WT mice were collected at d 7 and d 14 after MI,respectively,to verify the expression of myofibroblast markers(n=3).Results Compared with WT mice,LVEF and LVFS in adult Mydgf-KO mice showed no significant changes(all P＞0.05).However,the mRNA levels of α-SMA and postn were upregulated,and α-SMA protein expression was also increased(all P＜0.05).After MI,compared with WT mice,LVEF and LVFS in Mydgf-KO mice decreased,and the infarcted area increased significantly(all P＜0.05).Furthermore,mRNA levels of α-SMA,col8al,postn,and ctgf were increased in Mydgf-KO mice.In addition,the α-SMA protein expression level was upregulated and α-SMA-positive fibroblasts were increased(P＜0.05).Conclusion Mydgf deletion promotes the transformation of cardiac fibroblasts into myofibroblasts and aggravates myocardial fibrosis after MI.

Objective:To explore the effect of systematic diagnosis and treatment model based on new classification on primary hypospadias.Methods:The data of 689 patients with primary hypospadias admitted to Renji Hospital Affiliated to Shanghai Jiaotong University School of Medicine from March 2010 to June 2021 were retrospectively analyzed. Of all, 349 cases from March 2010 to June 2016 were treated with traditional treatment methods, and 340 cases from July 2016 to June 2021 were treated with systematic diagnosis and treatment model based on new hypospadias classification. Traditional treatment group of patients according to the European Society of Urology (EAU) guide classification, were divided into the distal-anterior type (located on the glans or distal shaft of the penis), intermediate-middle type (penile) and proximal-posterior type (penoscrotal, scrotal, perineal). For distal type, tubularized incised plate (TIP) or dorsal inlay urethroplasty (Inlay) was performed. Patients with intermediate-middle type underwent Inlay, and patients with proximal-posterior type underwent Bracka staging. In the systematic diagnosis and treatment group, hypospadias was reclassified into distal type (located on the glan or in the first third of the shaft of the penis), junctional type (the back two-thirds of the shaft of the penis and the base of the penis), and proximal type (scrotum or perineum). For distal type, TIP or Inlay was used. Patients with junctional type were treated with urethroplasty with union free graft and local flap method (Montage). Patients with proximal type were treated with Montage or Bracka staging. Patients with testicular volume <0.8 ml and glans width <1.0 cm were treated with human chorionic gonadotropin (HCG) before surgery. There were 349 patients in the traditional group, with an average age of (18.58±7.59) months. There were 157 cases of distal-anterior type, 47 cases of intermediate-middle type and 145 cases of proximal-posterior type. There were 181 cases with penile curvature < 30° and 168 cases with penile curvature ≥30°. There were 340 cases in the systematic diagnosis and treatment group, with an average age of (18.94±6.05) months. According to EAU classification, 160 cases were distal-anterior type, 42 cases were intermediate-middle type and 138 cases were proximal-posterior type. There were 197 cases with penile curvature < 30° and 143 cases with penile curvature ≥30°. There was no significant difference in the above indicators between the two groups ( P>0.05). The surgical methods, incidence of postoperative complications and the time of hospital stay between the two groups compared. The changes in penis size and testis volume before and after the use of hormones in the systematic treatment group were compared. Results:In the traditional group, 139 patients underwent TIP, 65 patients underwent Inlay, and 145 patients underwent Bracka staging. In the systematic diagnosis and treatment group, 187 cases were reclassified as distal type (153 cases underwent TIP and 34 cases underwent Inlay). Of the 69 cases of junctional type, 10 cases underwent Inlay, and 59 cases underwent Montage. Of the 84 cases of proximal type, 77 cases underwent Montage, and 7 cases underwent Bracka staging. There was significant difference between the two groups ( P<0.01). In the traditional group, there were 31 cases of urinary fistula (8.9%), 43 cases of urethral stricture (12.3%), 21 cases of urethral diverticulum (6.1%), 45 cases of penile recurvature (12.9%), and the total incidence of complications was 40.1% (140/349). There were 19 cases of urinary fistula (5.6%), 28 cases of urethral stricture (8.2%), 15 cases of urethral diverticulum (4.4%) and 24 cases of penile recurvature (7.1%) in the systematic diagnosis and treatment group. The total incidence of complications was 25.3%(86/340). There were statistically significant differences in the incidence of the above complications between the two groups ( P<0.05). There was a significant difference in the duration of hospital stay between the traditional group and the systematic diagnosis and treatment group [(4.3±1.2) d vs. (1.5±0.5) d, P=0.01]. The width of glans penis after HCG treatment was (1.35±0.14) cm, which was significantly higher than that before HCG treatment [(0.96±0.24) cm, P=0.03]. The length of penis [(2.55±0.19) cm vs. (2.29±0.16) cm] and the volume of testis [(0.76±0.24) ml vs. (0.64±0.15) ml] were not significantly different from those before treatment ( P>0.05). Conclusions:The new classification has clarified the choice of surgical methods, avoiding the amplification of surgical indications. The systematic diagnosis and treatment model based on new classification has significantly improved the treatment effect of hypospadias and effectively reduced postoperative complications. It provides an optional diagnosis and treatment model for hypospadias.

Objective:To evaluate postoperative calcitonin level as a prognostic marker in long-term follow-up of medullary thyroid carcinoma(MTC).Methods:Clinical data of 146 MTC cases treated at Tianjin Medical University Cancer Institute and Hospital from Jan 2011 to Dec 2019 were reviewed retrospectively. The relationship between postoperative calcitonin and disease-free survival was analyzed. According to the level of calcitonin six months after operation, patients were divided into normal level group and elevated group.Results:The median tumor size in those 146 cases was (1.78±1.22)cm, and 81 cases had lymph node metastasis. After 6 months of follow-up, 89 cases had normal calcitonin, with median tumor size of (1.63±1.20)cm, and 35 cases had lymph node metastasis . After a median follow-up of 56 months, 78 patients had normal calcitonin, 11 patients had biochemical relapse, 3 patients had structural relapse, and no patients died. 57 cases had a higher calcitonin ,median tumor size (1.97±1.22)cm, 46 cases had lymph node metastasis, 5 cases had distant metastasis, 18 cases had structural recurrence, and 7 patients died. Univariate analysis showed that lymph node metastasis, TNM stage, preoperative calcitonin, lymph node dissection and postoperative calcitonin were correlated with long-term disease-free survival (all P < 0.05). Multivariate analysis showed that postoperative calcitonin and TNM stage were an independent prognosis factor for disease-free survival in MTC patients (all P < 0.05). Conclusion:Postoperative calcitonin is a independent prognostic marker for long-term disease-free survival in MTC patients.

Objective:To evaluate the clinical and ultrasonographic features for early diagnosis and prediction of lateral cervical lymph node metastasis of medullary thyroid microcarcinoma.Methods:From Jan 2010 to Jan 2020 233 patients undergoing primary surgery were categorized as "medullary thyroid microcarcinoma" and "medullary thyroid macrocarcinoma". The preoperative clinical and ultrasonographic characteristics, the diagnostic positive rate of preoperative serum calcitonin and fine needle aspiration (FNA) were investigated between two groups. All patients with medullary thyroid microcarcinoma were divided on the basis of wether there was lateral cervical lymph node metastasis.Results:There were statistically significant differences in initial diagnostic method(χ 2=32.290, P=0.000), TNM staging(χ 2=50.300, P=0.000) between medullary thyroid microcarcinoma and medullary thyroid macrocarcinoma. Medullary thyroid microcarcinoma showed more malignant ultrasonic features. The diagnostic accuracy of preoperative serum calcitonin was higher than FNA for medullary thyroid micro carcinoma(χ 2=47.933, P=0.000). Multivariate regression analysis demonstrated that the abutment/perimeter ≥1/4( OR=25.475, 95%CI: 2.320-279.771), preoperative serum calcitonin >65 ng/L( OR=32.663, 95%CI:2.433-438.409) were the independent factor for lateral cervical lymph node metastases of medullary thyroid microcarcinoma. Conclusions:The combination of ultrasonography and serum calcitonin helps establish early diagnosis of medullary thyroid microcarcinoma. Medullary thyroid microcarcinoma with the abutment/perimeter ≥1/4, serum calcitonin >65 ng/L predicts lateral cervical lymph node metastases.

Objective: To study the clinical significance of serum calcitonin in the diagnosis and treatment of medullary thyroid carcinoma and to analyze its cost-benefit. Methods: One hundred and forty one patients with medullary thyroid carcinoma who undertook calcitonin test and frozen pathological examination were enrolled in this study from Oct 2012 to Mar 2018. Using the method of χ² test, the positive rate of calcitonin test and frozen pathological examination in diagnosis of medullary thyroid carcinoma(MTC) were compared. Firstly, we compared the correct checkout cost of calcitonin test and that of frozen pathological examination (total number of patients×cost of examination/the correctly detected number of patients) . Secondly, we calculated whether calcitonin test help patients save money(average cost of treatment in hospital for MTC×number of patients who were evaluated to be candidate for surgery-cost of calcitonin test×total number of patients)/total number of patients. Results: 139 patients were positive in calcitonin test among 141 patients, and the positive rate was 98.58%. 91 patients were positive in frozen pathological examination, and the positive rate was 64.54% (χ²=97.821, P<0.000 1) . Cost-benefit analysis showed that the correct checkout cost of calcitonin test and frozen pathological examination were 71.01 yuan and 426.10 yuan, also,1 371 938.64 yuan could be saved totally and 9 730.06 yuan could be saved per patient because of calcitonin test. Conclusion Serum calcitonin test had a significant effect on the diagnosis and treatment of medullary thyroid carcinoma and was economical and practical.