ObjectiveTo evaluate the clinical efficacy and safety of Jingmaiyan granules （composed of Lonicerae Japonicae Flos， Sedi Herba， Paeoniae Radix Rubra， Moutan Cortex， Rhei Radix et Rhizoma Praeparata， and Glycyrrhizae Radix et Rhizoma） combined with external application of Jinhuang Ointment in treating acute-stage blood heat stasis type superficial thrombophlebitis （ST） of lower extremities， and to explore their effects on hemorheology and serum inflammatory factors. MethodsA randomized， double-blind， placebo-controlled clinical trial was conducted. A total of 124 patients with lower extremity ST were randomized into two groups（62 cases in each group）. The control group received external application of Jinhuang ointment and oral placebo treatment， while the observation group received external application of Jinhuang ointment and oral Jingmaiyan granules. Both groups were treated for 2 weeks. The clinical symptom scores， therapeutic efficacy of traditional Chinese medicine （TCM） syndrome， pain visual analog scale （VAS） scores， hemorheological indices ［including whole blood high-shear， medium-shear， and low-shear viscosity， as well as plasma viscosity （PV）］， and inflammatory factors ［C-reactive protein （CRP）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）］ were compared before and after treatment. ResultsAfter 2 weeks of treatment， the total effective rate in the observation group （98.3%， 60/62） was significantly higher than that in the control group （83.8%， 52/62）， with a statistically significant difference （Z=3.512 1， P<0.05）. Compared with pre-treatment scores， the scores for skin color， skin temperature， swelling， pain， and cord or nodules were significantly reduced in both groups （P<0.05）， with more pronounced improvement in the observation group （P<0.05）. Additionally， compared with pre-treatment levels， the whole blood viscosity （low-， medium-， and high-shear） significantly improved in both groups after treatment （P<0.05）， with more marked improvement in the observation group （P<0.05）. Furthermore， the plasma viscosity， CRP， IL-6， and TNF-α levels were significantly reduced in both groups after treatment （P<0.05）， with more pronounced improvement observed in the observation group （P<0.05）. ConclusionThe combination of external application of Jinhuang ointment and oral Jingmaiyan granules effectively improves clinical symptoms， hemorheological abnormalities， and inflammatory responses in patients with acute stage blood heat stasis type ST of lower extremities. The treatment is safe and holds clinical promotion value.

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

ObjectiveTo observe the effects of the Fuyuan Tongluo prescription （composed of Astragali Radix， Carthami Flos， Spatholobi Caulis， Liquidambaris Fructus， Lycopodii Herba， Centellae Herba， etc.） in the treatment of restenosis with collateral obstruction syndrome after interventional operation of lower limb arteriosclerosis obliterans， and its impact on the primary patency rate. MethodsA total of 88 patients with collateral obstruction syndrome after interventional surgery for lower limb arteriosclerosis obliterans were randomly divided into two groups. The control group （n₁=44） received dual antiplatelet therapy with aspirin and clopidogrel. The observation group （n₂=44） was treated with Fuyuan Tongluo prescription non-decocted granules in addition to aspirin and clopidogrel. Both groups received treatment for 24 weeks and were followed up for 36 weeks. The changes in primary patency rate， symptom scores， ankle-brachial index （ABI）， coagulation function， and inflammatory markers before and after treatment were compared between the two groups. ResultsFor primary patency rate， after 36 weeks of treatment， the observation group had a significantly better primary patency rate than the control group （χ²=4.14，P<0.05）. After 24 weeks of treatment， there was no significant difference in primary patency rate between the two groups. Clinical efficacy comparison： Based on symptom quantification scores， and using the Nimodipine method as a reference， the overall efficacy of the observation group was superior to that of the control group after 24 weeks of treatment （χ²=2.24，P<0.05）. ABI levels： The observation group had a higher ABI than the control group after 24 and 36 weeks of treatment （P<0.05）. Coagulation function indicators： After 24 and 36 weeks of treatment， D-dimer and fibrinogen levels in both groups were lower than before treatment （P<0.05）. Inflammatory markers： After 24 and 36 weeks of treatment， CRP levels in the observation group were lower than those in the control group （P<0.05）. There were no significant differences in white blood cell （WBC） and erythrocyte sedimentation rate （ESR） levels before and after treatment between the two groups. ConclusionAdding Fuyuan Tongluo prescription non-decocted granules to dual antiplatelet therapy can improve the primary patency rate of the affected vessels in patients with lower limb arteriosclerosis obliterans after interventional surgery. Longer use of Fuyuan Tongluo prescription can significantly improve clinical symptoms， demonstrating clinical application value.

ObjectiveTo observe the effects of the Fuyuan Tongluo prescription （composed of Astragali Radix， Carthami Flos， Spatholobi Caulis， Liquidambaris Fructus， Lycopodii Herba， Centellae Herba， etc.） in the treatment of restenosis with collateral obstruction syndrome after interventional operation of lower limb arteriosclerosis obliterans， and its impact on the primary patency rate. MethodsA total of 88 patients with collateral obstruction syndrome after interventional surgery for lower limb arteriosclerosis obliterans were randomly divided into two groups. The control group （n₁=44） received dual antiplatelet therapy with aspirin and clopidogrel. The observation group （n₂=44） was treated with Fuyuan Tongluo prescription non-decocted granules in addition to aspirin and clopidogrel. Both groups received treatment for 24 weeks and were followed up for 36 weeks. The changes in primary patency rate， symptom scores， ankle-brachial index （ABI）， coagulation function， and inflammatory markers before and after treatment were compared between the two groups. ResultsFor primary patency rate， after 36 weeks of treatment， the observation group had a significantly better primary patency rate than the control group （χ²=4.14，P<0.05）. After 24 weeks of treatment， there was no significant difference in primary patency rate between the two groups. Clinical efficacy comparison： Based on symptom quantification scores， and using the Nimodipine method as a reference， the overall efficacy of the observation group was superior to that of the control group after 24 weeks of treatment （χ²=2.24，P<0.05）. ABI levels： The observation group had a higher ABI than the control group after 24 and 36 weeks of treatment （P<0.05）. Coagulation function indicators： After 24 and 36 weeks of treatment， D-dimer and fibrinogen levels in both groups were lower than before treatment （P<0.05）. Inflammatory markers： After 24 and 36 weeks of treatment， CRP levels in the observation group were lower than those in the control group （P<0.05）. There were no significant differences in white blood cell （WBC） and erythrocyte sedimentation rate （ESR） levels before and after treatment between the two groups. ConclusionAdding Fuyuan Tongluo prescription non-decocted granules to dual antiplatelet therapy can improve the primary patency rate of the affected vessels in patients with lower limb arteriosclerosis obliterans after interventional surgery. Longer use of Fuyuan Tongluo prescription can significantly improve clinical symptoms， demonstrating clinical application value.

ObjectiveTo investigate the effect and mechanism of Yishen Tongluo prescription in protecting mice from oxidative stress injury in diabetic kidney disease （DKD） via the nuclear factor erythroid 2-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1）/NAD（P）H quinone oxidoreductase 1 （NQO1） signaling pathway. MethodsSpecific pathogen-free （SPF） male C57BL/6 mice were assigned into a control group （n=10） and a modeling group （n=50）. The DKD model was established by intraperitoneal injection of streptozotocin. The mice in the modeling group were randomized into a model group， a semaglutide （40 μg·kg^-1） group， and high-， medium-， and low-dose （18.2， 9.1， 4.55 g·kg^-1， respectively） Yishen Tongluo prescription groups， with 10 mice in each group. The treatment lasted for 12 weeks. Blood glucose and 24-h urine protein levels were measured， and the kidney index （KI） was calculated. Serum levels of creatinine （SCr）， blood urea nitrogen （BUN）， alanine aminotransferase （ALT）， and aspartate aminotransferase （AST） were assessed. The pathological changes in the renal tissue were evaluated by hematoxylin-eosin， periodic acid-Schiff， periodic acid-silver methenamine， and Masson’s trichrome staining. Enzyme-linked immunosorbent assay kits were used to measure the levels of β2-microglobulin （β2-MG）， neutrophil gelatinase-associated lipocalin （NGAL）， kidney injury molecule-1 （KIM-1）， liver fatty acid-binding protein （L-FABP）， nitric oxide synthase （NOS）， glutathione （GSH）， total antioxidant capacity （T-AOC）， and 8-hydroxy-2'-deoxyguanosine （8-OHdG）. Immunohistochemical staining was performed to examine the expression of Kelch-like ECH-associated protein 1 （Keap1） and malondialdehyde （MDA）. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR） and Western blot were employed to determine the mRNA and protein levels， respectively， of factors in the Nrf2/HO-1/NQO1 signaling pathway. ResultsCompared with the control group， the DKD model group showed rises in blood glucose， 24-h urine protein， KI， SCr， BUN， and ALT levels， along with glomerular hypertrophy， renal tubular dilation， thickened basement membrane， mesangial expansion， and collagen deposition. Additionally， the model group showed elevated levels of β2-MG， NGAL， KIM-1， L-FABP， NOS， and 8-OHdG， lowered levels of GSH and T-AOC， up-regulated expression of MDA and Keap1， and down-regulated expression of Nrf2， HO-1， NQO1， and glutamate-cysteine ligase catalytic subunit （GCLC） （P<0.05）. Compared with the model group， the semaglutide group and the medium- and high-dose Yishen Tongluo prescription groups showed reductions in blood glucose， 24-h urine protein， KI， SCr， BUN， and ALT levels， along with alleviated pathological injuries in the renal tissue. In addition， the three groups showed lowered levels of β2-MG， NGAL， KIM-1， L-FABP， NOS， and 8-OHdG， elevated levels of GSH and T-AOC， down-regulated expression of MDA and Keap1， and up-regulated expression of Nrf2， HO-1， NQO1， and GCLC （P<0.05）. ConclusionYishen Tongluo prescription exerts renoprotective effects in the mouse model of DKD by modulating the Nrf2/HO-1/NQO1 signaling pathway， mitigating oxidative stress， and reducing renal tubular injuries.

Objective: To investigate the mechanism of Yishen Tongluo Formula in ameliorating renal pyroptosis in diabetic nephropathy mice by regulating the toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor-κB (NF-κB) signaling pathway. Methods: Sixty C57BL/6 male mice were randomly divided into control (10 mice) and intervention groups (50 mice) using random number table method. The diabetes nephropathy model was established by intraperitoneally injecting streptozotocin(50 mg/kg). After modeling, the intervention group was further divided into model, semaglutide (40 μg/kg), and high-, medium-, and low-dose Yishen Tongluo Formula groups (15.6, 7.8, and 3.9 g/kg, respectively) using random number table method. The high-, medium-, and low-dose Yishen Tongluo Formula groups were administered corresponding doses of medication by gavage, the semaglutide group received a subcutaneous injection of semaglutide injection, and the control group and model groups were administered distilled water by gavage for 12 consecutive weeks. Random blood glucose levels of mice in each group were monitored, and the 24-h urinary protein content was measured using biochemical method every 4 weeks; after treatment, the serum creatinine and urea nitrogen levels were measured using biochemical method. The weight of the kidneys was measured, and the renal index was calculated. Hematoxylin and eosin, periodic acid-Schiff, periodic Schiff-methenamine, and Masson staining were used to observe the pathological changes in renal tissue. An enzyme-linked immunosorbent assay was used to detect urinary β2-microglobulin (β2-MG), neutrophil gelatinase-associated lipocalin (NGAL), and kidney injury molecule-1 (KIM-1) levels. Western blotting and real-time fluorescence PCR were used to detect the relative protein and mRNA expression levels of nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3), Caspase-1, gasdermin D (GSDMD), interleukin-1β (IL-1β), and interleukin-18 (IL-18) in renal tissue. Immunohistochemistry was used to detect the proportion of protein staining area of the TLR4, MyD88, and NF-κB in renal tissue. Results: Compared with the control group, the random blood glucose, 24-h urinary protein, serum creatinine, urea nitrogen, and renal index of the model group increased, and the urine β2-MG, NGAL, and KIM-1 levels increased. The relative protein and mRNA expression levels of NLRP3, Caspase-1, GSDMD, IL-1β, and IL-18 in renal tissue increased, and the proportion of TLR4, MyD88, and NF-κB protein positive staining areas increased (P<0.05). Pathological changes such as glomerular hypertrophy were observed in the renal tissue of the model group. Compared with the model group, the Yishen Tongluo Formula high-dose group showed a decrease in random blood glucose after 12 weeks of treatment (P<0.05). The Yishen Tongluo Formula high- and medium-dose groups showed a decrease in 24-h urinary protein, creatinine, urea nitrogen, and renal index, as well as decreased β2-MG, NGAL, and KIM-1 levels. NLRP3, Caspase-1, GSDMD, IL-1 β, and IL-18 relative protein and mRNA expression levels were also reduced, and the proportion of TLR4, MyD88, and NF-κB protein positive staining areas was reduced (P<0.05). Pathological damage to renal tissue was ameliorated. Conclusion Yishen Tongluo Formula may exert protective renal effects by inhibiting renal pyroptosis and alleviating tubular interstitial injury in diabetic nephropathy mice by regulating the TLR4/MyD88/NF-κB signaling pathway.

A porous composite scaffold is prepared through freeze-drying method using sodium alginate as the matrix and calcium citrate double salt as the functional enhancer.To thoroughly investigate the structure and properties of the composite scaffold,advanced instruments including Fourier transform infrared spectrometer,X-ray diffractometer,scanning electron microscope,and thermogravimetric analyzer are utilized for comprehensive structural characterization.Meanwhile,key properties such as porosity,mechanical performance,swelling ratio,gel fraction,and in vitro degradation rate are systematically tested and analyzed.The results show that the composite of sodium alginate and calcium citrate double salt is a physical complex.The porosity of the scaffold mainly ranges from 200 to 600 μm,and as the addition of double salt increases,the pore size firstly increases and then slightly decreases,while the pore distribution become more uneven.In terms of mechanical properties,with the gradual increase of the content of double salt in the scaffold,the mechanical properties of the scaffold are significantly improved.However,its swelling ratio and gel fraction exhibit decreasing trends.Additionally,the degradation rate of the alginate scaffold is relatively fast,while the addition of calcium citrate double salt does not significantly improve the degradation rate.This study not only provides valuable insights into the preparation and property research of porous composite scaffold composed of sodium alginate/calcium citrate double salt,but also offers important reference for their potential applications in biomedicine.

A porous composite scaffold is prepared through freeze-drying method using sodium alginate as the matrix and calcium citrate double salt as the functional enhancer.To thoroughly investigate the structure and properties of the composite scaffold,advanced instruments including Fourier transform infrared spectrometer,X-ray diffractometer,scanning electron microscope,and thermogravimetric analyzer are utilized for comprehensive structural characterization.Meanwhile,key properties such as porosity,mechanical performance,swelling ratio,gel fraction,and in vitro degradation rate are systematically tested and analyzed.The results show that the composite of sodium alginate and calcium citrate double salt is a physical complex.The porosity of the scaffold mainly ranges from 200 to 600 μm,and as the addition of double salt increases,the pore size firstly increases and then slightly decreases,while the pore distribution become more uneven.In terms of mechanical properties,with the gradual increase of the content of double salt in the scaffold,the mechanical properties of the scaffold are significantly improved.However,its swelling ratio and gel fraction exhibit decreasing trends.Additionally,the degradation rate of the alginate scaffold is relatively fast,while the addition of calcium citrate double salt does not significantly improve the degradation rate.This study not only provides valuable insights into the preparation and property research of porous composite scaffold composed of sodium alginate/calcium citrate double salt,but also offers important reference for their potential applications in biomedicine.

OBJECTIVES: Diabetes mellitus is closely associated with sudden sensorineural hearing loss (SSNHL), but no definitive evidence has established a causal relationship between type 1 diabetes mellitus (T1DM) and SSNHL. This study aims to investigate the impact of T1DM on SSNHL from a genetic perspective, providing insights for risk prediction and treatment strategies. METHODS: Genetic data related to exposure (T1DM) and outcome (SSNHL) were obtained from publicly available genome-wide association studies (GWAS). Instrumental variables were selected, and Mendelian randomization (MR) analysis was conducted to explore the causal association between T1DM and SSNHL. Inverse variance weighted (IVW) analysis was used as the primary method, with random-effects IVW serving as the main analytical approach. MR-Egger, weighted median, simple mode, and weighted mode analyses were utilized as supplementary methods. Cochran's Q test was applied to evaluate the heterogeneity of the selected instrumental variables, MR-PRESSO was applied to detect outliers, MR-Egger regression was used to assess horizontal pleiotropy and leave-one-out analysis was conducted to examine the robustness of individual single nucleotide polymorphisms (SNPs) on the overall results. RESULTS: A total of 127 SNPs were selected as instrumental variables for the MR analysis. IVW analysis demonstrated a genetically determined association between T1DM and SSNHL (OR=1.036, 95% CI 1.002 to 1.071, P=0.038). Forest plots and scatter plots indicated a causal relationship, suggesting that T1DM increases the risk of SSNHL. Cochran's Q test demonstrated no significant heterogeneity among SNPs (MR-Egger: Q=126.030, P=0.356; IVW: Q=126.450, P=0.373). The funnel plot appeared symmetrical, indicating that the selected instrumental variables were primarily related to exposure rather than potential confounding factors. The MR-Egger intercept was not significantly different from zero (P=0.527), indicating no evidence of horizontal pleiotropy among the SNPs. MR-PRESSO analysis did not identify any outlier SNPs (P=0.356). Leave-one-out analysis confirmed the robustness of the findings, as the results remained stable after removing individual SNPs. CONCLUSIONS Two-sample MR analysis supports the conclusion that T1DM patients have an increased risk of developing SSNHL.
Humans ; Mendelian Randomization Analysis ; Hearing Loss, Sensorineural/etiology* ; Diabetes Mellitus, Type 1/genetics* ; Genome-Wide Association Study ; Hearing Loss, Sudden/etiology* ; Polymorphism, Single Nucleotide ; Risk Factors ; Genetic Predisposition to Disease

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.