This study aims to establish a high-performance size-exclusion chromatography (HPSEC) method for determining the content of the classical swine fever virus (CSFV) E2 protein and screen the optimal stabilizer to enhance the stability of this protein. The optimal detection conditions were determined by optimizing the composition of the mobile phase, and characteristic chromatographic peaks were identified by SDS-PAGE and Western blotting. The specificity, repeatability, precision, linearity, limit of detection (LOD), and limit of quantitation (LOQ) of the method were assessed. The method established was used to determine the content of CSFV E2 protein antigen and vaccine. Differential scanning fluorimetry (DSF) was employed to screen the buffer system, pH, and salt ion concentrations, and sugar, amino acid, and alcohol stabilizers were further screened. The results showed that using a 200 mmol/L phosphate buffer provided the best column efficiency. An antigen-specific chromatographic peak appeared at the retention time of 18 min, which was identified as the CSFV E2 protein by SDS-PAGE and Western blotting. The method exhibited high specificity for detecting the CSFV E2 protein, with no absorbance peak observed in the blank control. The relative standard deviation (RSD) of the peak area for six repeated injections of the CSFV E2 protein was 0.74%, indicating good repeatability of the method. The RSD for repeated detection of two different concentrations of CSFV E2 protein samples by different operators at different time points was less than 2%, suggesting good intermediate precision of the method. The peak area of the CSFV E2 protein was linearly related to its concentration, with the regression equation showing R² of 1.000. The LOD and LOQ of the method were 14.88 μg/mL and 29.75 μg/mL, respectively. Application of the developed method in the detection of three batches of CSFV E2 protein antigen and three batches of vaccine demonstrated results consistent with those from the bicinchoninic acid (BCA) assay, which meant that the method could accurately determine the content of CSFV E2 protein antigen and vaccine. The DSF method identified 50 mmol/L Tris-HCl at pH 8.0 as the optimal buffer, and the addition of sugar and alcohol stabilizers further improved the stability of the CSFV E2 protein. The HPSEC method established in this study is simple, fast, and exhibits good accuracy and repeatability, enabling precise measurement of the CSFV E2 protein content. It is expected to play a crucial role in the quality control of the CSFV E2 vaccine. Furthermore, the strategy for improving the CSFV E2 protein stability, identified through DSF screening, has significant implications for enhancing the stability of the CSFV E2 vaccine.
Classical Swine Fever Virus/chemistry* ; Chromatography, Gel/methods* ; Animals ; Swine ; Viral Envelope Proteins/immunology*

Malacoplakia is a rare granuloma disease mainly occurred in the urinary system, it is even rarer for renal malacoplakia invading the descending colon complicated with bladder malacoplakia. In this study, one such case was reported. Imaging examination suggested that the left kidney was a large patchy mixed density shadow, and enhancement scan lesion was uneven enhancement. CT guided renal puncture biopsy was performed, and postoperative pathology suggested renal malacoplakia. Transurethral cystoscopy was performed, and postoperative pathology confirmed that it was malacoplakia of the bladder. The effect of conservative antibiotic treatment was not good. The patient underwent radical nephrectomy + left hemicolectomy under general anesthesia, and postoperative pathology confirmed the diagnosis of renal malacoplakia, which involved the mucosa of the intestinal tube and the entire muscular layer. The patient was followed up for 6 months after surgery, and no recurrence was seen on CT.

Hemangioma of the penile head is rare. This paper reported a patient, 16 years old, who was admitted to hospital due to the discovery of multiple masses on the head of the penis for more than two years. Physical examination showed that three vascular mass-like masses were distributed along the coronal sulcus at the 3, 9, and 12 points of the penile head, and the larger one was about 10 mm×5 mm size, blue-purple, soft, and painless. Ultrasound examination suggested that the patient had a penile head hemangioma. Surgical resection was performed, and the postoperative pathological diagnosis was penile head hemangioma.The follow-up of 3 months showed that the wound healed well without recurrence, and the penile head appearance was not obviously deform.

Objective:To compare the efficacy and safety of programmed death-1(PD-1)inhibitors in combination with chemotherapy versus chemotherapy alone in patients with metastatic bladder cancer.Methods:A retrospective analysis was performed on the clinical data of 77 cases of metastatic bladder cancer who were admitted to the First Affiliated Hospital of Zhengzhou University from January 2014 to October 2022. According to the different clinical treatment regimens, they were divided into two groups. Patients treated with PD-1 inhibitors combined with gemcitabine and cisplatin (GC) regimen were referred to as IGC group, and patients who received GC chemotherapy alone were referred to as GC group. There were 24 cases in IGC group, including 18 males and 6 females, with a median age of 60 (56, 67) years old. Seventeen cases had a history of smoking. Fifteen cases had an Eastern Cooperative Oncology Group (ECOG) score of 0 and 9 cases had a score of 1. Twenty-three cases suffered distant metastasis (stage M 1). Two cases suffered lymph node metastasis only, 6 cases with liver metastasis, 8 cases with lung metastasis, and 8 cases with bone metastasis. There were 53 cases in GC group, including 45 males and 8 females, with a median age of 63 (55, 69) years old. Thirty-one cases had a history of smoking. Thirty cases had an ECOG score of 0 and 23 cases had a score of 1, 48 cases suffered stage M 1, 2 cases suffered lymph node metastasis only. Nineteen cases suffered liver metastasis. Twenty-seven cases suffered lung metastasis. And 23 cases suffered bone metastasis. There was no statistically significant difference in the above general information between the two groups ( P>0.05). Kaplan-Meier method was used to plot the survival curves, and the difference of median progression-free survival (mPFS) and median overall survival (mOS) between the two groups was compared by log-rank test. Finally, the difference in adverse reactions between the two groups was compared. Results:The objective response rate (ORR) was 41.7% and the disease control rate (DCR) was 87.5% in the IGC group.As a comparison, the ORR was 43.4% and the DCR was 83.0% in the GC group. The differences in ORR ( P=0.887) and DCR ( P=0.871) between the two groups were not statistically significant. All patients were followed up for 3 to 45 months, with a median follow-up time of 24 (14, 43) months. The mPFS was 7.0 (95% CI 5.7-8.3) months in the GC group and 8.0 (95% CI 3.0-13.1) months in the IGC group, and the difference was statistically significant between the two groups ( P=0.026). The mOS of patients in the GC group was 16.0 (95% CI 14.4-17.6) months, the mOS was not yet reached in the IGC group, and patients in the IGC group had longer mOS with a statistically significant difference ( P=0.022). All patients experienced treatment-related adverse reactions. Grade 3-4 adverse reactions occurred in 8 cases (33.3%) in the IGC group and in 16 cases (30.2%) in the GC group, and no adverse reaction-related deaths were observed( P=0.992). The most common adverse reactions in both groups were anemia, including 18 cases (75.0%) in IGC group and 38 cases (71.7%) in GC group. There were 4 cases (16.7%) of grade 1-2 hypothyroidism in the IGC group but no patients with hypothyroidism were found in the GC group, and the difference was statistically significant ( P=0.012). There were 8 cases (33.3%) and 4 cases (7.5%) of grade 1-2 skin adverse reactions in the IGC and GC groups, respectively, and the difference was statistically significant ( P=0.011). The immune-related adverse reactions of PD-1 inhibitors in IGC group were 1 case of hyperthyroidism (4.2%), 4 cases of hypothyroidism (16.7%), 1 case of adrenal insufficiency (4.2%), and 1 case of immune colitis (4.2%). Conclusions:Compared with chemotherapy alone, PD-1 inhibitors combined with chemotherapy for metastatic bladder cancer can effectively prolong the mPFS and median mOS. The adverse reactions of the two groups were tolerable, and there was no significant difference in the incidence of grade 3-4 adverse reactions. In general, PD-1 inhibitors combined with chemotherapy in the treatment of metastatic bladder cancer is safe and feasible, but attention should be paid to the immune-related adverse reactions of PD-1 inhibitors.

Objective:To investigate the clinical characteristics and prognostic factors of primary urethral cancer.Methods:The clinical data of 35 patients with primary urethral cancer admitted to the First Affiliated Hospital of Zhengzhou University from January 2011 to April 2022 were retrospectively analyzed. There were 12 males (34.3%) and 23 females (65.7%). The average age was 61.1 ± 13.0 years old. The clinical symptoms included 13 cases of urethral obstruction (37.1%), 7 cases of hematuria (20.0%), 6 cases of urethral bleeding (17.1%), 5 cases of urinary tract irritation (14.3%), 1 case of Urinary incontinence (2.9%), 1 case of low back pain (2.9%), 1 case of scrotal ulcer (2.9%), and 1 case (2.9%) by self examination. All patients underwent cystourethroscopy and tissue biopsy. The biopsy pathology showed 16 cases of urothelial carcinoma, 7 cases of squamous carcinoma, 4 cases of adenocarcinoma, 3 cases of malignant melanoma, 1 case of urothelial carcinoma with squamous carcinoma, 1 case of Signet ring cell carcinoma, 1 case of sarcomatoid carcinoma, 1 case of embryonic Rhabdomyosarcoma, and 1 case of epithelioid angiosarcoma. The tumors were located in the proximal urethra in 13 cases (37.1%) and in the distal urethra in 22 cases (62.9%). There were 14 cases (40.0%) with a maximum diameter of less than 3 cm, 16 cases (45.7%) with a diameter of ≥ 3 cm, and 5 cases (14.3%) with mucosal abnormalities. There were 12 cases of T 1 stage, 9 cases of T 2 stage, 7 cases of T 3 stage, and 7 cases of T 4 stage in tumor staging. Imaging evaluation of lymph nodes showed 25 cases of N 0 stage, 2 cases of N 1 stage, and 8 cases of N 2 stage; A total of 11 cases of lymph node biopsy were performed (including 8 cases of intraoperative lymph node dissection and 3 cases of preoperative lymph node biopsy), of which 6 cases had lymph node metastasis, and 1 case was initially diagnosed with distant metastasis. Thirty-one cases underwent surgical treatment, of which 16 cases underwent radical urethrectomy, and 8 cases underwent intraoperative pelvic and bilateral inguinal lymph node dissection, 8 cases underwent resection of urethral tumors, and 7 cases underwent transurethral resection of tumors. Four cases did not undergo surgical treatment, while 1 case had epithelioid angiosarcoma and received radiotherapy combined with chemotherapy, 2 cases received chemotherapy with GC (Gemcitabine+ cisplatin) regimen, and 1 case received immunotherapy with immune checkpoint inhibitors. The risk factors that affected patient prognosis were analyzed. Results:All 35 cases in this group were followed up, with a median follow-up time of 22 (2, 122) months. Seventeen cases survived, 18 cases died, and the overall median survival duration was 23 (13 to not reached) months. The overall 5-year survival rate was 45%. The results of univariate analysis showed that clinical T-stage ( P=0.019), maximum tumor diameter ( P=0.016), and tumor location ( P=0.006) were independent risk factors affecting patient prognosis. Result of multivariate analysis showed that the maximum diameter of the tumor ≥ 3 cm ( HR=2.673, P=0.029) and the proximal location of the tumor ( HR=3.064, P=0.023) were independent risk factors affecting patient survival. Gender, age, treatment method, lymph node dissection, adjuvant radiotherapy, adjuvant chemotherapy, clinical manifestations, pathological type, clinical N staging, and pathological N staging had no significant impact on patient survival rate ( P>0.05). Single factor analysis was conducted on female patients separately, and only tumor location was found to be a prognostic factor ( χ2=17.246, P<0.01). Conclusions:Primary urethral cancer is a rare disease with various symptoms and poor prognosis. The maximum diameter of the tumor ≥3 cm and the tumor located at the proximal end of the urethra are clinical risk factors affecting the prognosis of patients with primary urethral cancer.

BACKGROUND: Nowadays, biological tissue engineering is a growing field of research. Biocompatibility is a key indicator for measuring tissue engineering biomaterials, which is of great significance for the replacement and repair of damaged tissues. METHODS: In this study, using gelatin, carboxymethyl chitosan, and sodium alginate, a tissue engineering material scaffold that can carry cells was successfully prepared. The material was characterized by Fourier transforms infrared spectroscopy. In addition, the prepared scaffolds have physicochemical properties, such as swelling ratio, biodegradability.we observed the biocompatibility of the hydrogel to different adult stem cells (BMSCs and ADSCs) in vivo and in vitro. Adult stem cells were planted on gelatin-carboxymethyl chitosan-sodium alginate (Gel/SA/CMCS) hydrogels for 7 days in vitro, and the survival of stem cells in vitro was observed by live/died staining. Gel/SA/CMCS hydrogels loaded with stem cells were subcutaneously transplanted into nude mice for 14 days of in vivo culture observation. The survival of adult stem cells was observed by staining for stem cell surface markers (CD29, CD90) and Ki67. RESULTS: The scaffolds had a microporous structure with an appropriate pore size (about 80 lm). Live/died staining showed that adult stem cells could stably survive in Gel/SA/CMCS hydrogels for at least 7 days. After 14 days of culture in nude mice, Ki67 staining showed that the stem cells supported by Gel/SA/CMCS hydrogel still had high proliferation activity. CONCLUSION Gel/SA/CMCSs hydrogel has a stable interpenetrating porous structure, suitable swelling performance and degradation rate, can promote and support the survival of adult stem cells in vivo and in vitro, and has good biocompatibility. Therefore, Gel/SA/CMCS hydrogel is a strong candidate for biological tissue engineering materials.

The clinical data of 5 patients with autoimmune encephalitis admitted to the psychiatric department of the 904th Hospital of the Joint Logistics Service Force from January 2016 to June 2020 were retrospectively analyzed. Among 5 patients, 4 had stress psychological events within one month before the onset, and 3 had precursor symptoms such as fever and vomiting. They were all characterized by rapid progress of atypical mental and behavioral abnormalities and cognitive impairment. In terms of neurological symptoms, 1 case had faciobrachial dystonic seizures (FBDS), 3 cases had seizures, 2 cases had involuntary movement, and 4 cases had autonomic dysfunction, including central hypopnea, arrhythmia, blood pressure instability and paroxysmal facial flushing. Most neurological symptoms occur within 1 month of the onset. MRI revealed abnormalities in cerebral cortex, thalamus, temporal lobe and insular lobe in 4 cases; EEG demonstrated bilateral short-range medium amplitude θ wave in 2 cases. Abnormal cerebrospinal fluid (CSF) pressure was detected in 4 cases and 2 cases had abnormal cell number CSF. Three patients had positive anti-N-methyl-D-aspartate receptor (NMDAR) antibody, one patient had positive anti-LGI1 antibody, and one patient had positive anti-γ-aminobutyric acid B receptor (GABA BR) antibody. One case was discharged automatically, the remaining 4 patients were treated with glucocorticoid or combined with gamma globulin and cyclophosphamide, antiepileptic drugs, antipsychotic drugs and other symptomatic treatment, and their symptoms were relieved. Patients were followed up for six months, there was slightly slow residual reaction in 2 cases and personality change in 1 case. Autoimmune encephalitis characterized by mental symptoms is likely to be misdiagnosed as mental disorders. Clinicians should identify symptoms different from mental disorders, taking into account of the possibility of autoimmune encephalitis, to make early diagnosis and treatment.

Objective:To study the relationship between white matter injury (WMI) and brain maturity in preterm infants at full-term corrected gestational age (cGA).Methods:A retrospective study was performed in preterm infants [GA≤32 weeks or birth weight (BW) ≤1 500 g] admitted to the neonatal intensive care unit of the First Affiliated Hospital of Xi'an Jiaotong University from January 2017 to August 2018 and the Northwest Women and Children's Hospital from January 2017 to June 2017. The infants received conventional magnetic resonance imaging (MRI) at cGA 37~42 weeks. The infants were assigned into the WMI group and the control group according to the WMI scoring system, including the total maturation scores (TMS) and four sub-item scores.Results:A total of 118 premature infants were enrolled in this study (17 cases in the WMI group and 101 cases in the control group). The GA was (30.3±1.7) weeks, and BW was (1 356±268) g. The proportion of delayed TMS in the WMI group was significantly higher than the control group [58.8%(10/17) vs. 31.7%(32/101), P<0.05]. The TMS of the WMI group were significantly lower than the control group [(10.7±1.8) vs. (11.8±1.5), P<0.05]. The sub-item scores of TMS showed that the myelination [(2.8±0.6) vs. (3.1±0.4), P<0.05] and glial cell migration bands of the WMI group [(1.6±0.4) vs. (2.1±0.6), P=0.004] were significantly lower than the control group and no significant differences existed in cortical folding and involution of germinal matrix tissue scores between the two groups. Conclusions:The brain maturity of preterm infants with WMI is substantially delayed than those without WMI, including delayed myelination and delayed disappearance of glial cell migration bands.

Objective:To explore the clinical features and prognosis of urachal carcinoma.Methods:The clinical data of 35 patients with urachal carcinoma admitted to the First Affiliated Hospital of Zhengzhou University from August 2011 to November 2019 were analyzed retrospectively. There were 23 males and 12 females, with a male to female ratio of 1.92∶1. The average age was (52.1±13.9) years old, and the median age was 55 years old. There were 8 patients with a history of smoking and 3 patients with a history of drinking. There were 5 cases of hypertension, 5 cases of diabetes, 2 cases of coronary heart disease, and 1 case of cerebral infarction. The first symptoms were hematuria in 25 cases, lower abdominal pain in 4 cases, abdominal mass in 2 cases, umbilical discharge in 1 case, and asymptomatic in 4 cases.Preoperative CT examination showed that the tumor was located on the top wall of the bladder in 24 cases and the front wall of the bladder in 11 cases. There were 25 solid tumors and 10 cystic tumors. The maximum diameter of the tumor was 1.5 to 11.0 cm, and the median maximum diameter of the tumor was 4.0 cm. Preoperative cystoscopy detected masses on the anterior or top wall of the bladder and urachal carcinoma was suspected in 35 cases. All 35 cases underwent enlarged partial cystectomy (conventional resection of the umbilical part), and 3 cases underwent pelvic lymph node dissection at the same time. Among them, 19 cases underwent open surgery, 14 cases underwent laparoscopic surgery, and 2 cases underwent Da Vinci robot assisted laparoscopic surgery.Results:According to Mayo staging, there were 10 cases of stage Ⅰ, 18 cases of stage Ⅱ, 1 case of stage Ⅲ, and 6 cases of stage Ⅳ. The overall follow-up rate was 91.4% (32/35), and the median follow-up time was 41 (3-103) months. The 1-year survival rate was 82.5%, the 3-year survival rate was 59.3%, and the 5-year survival rate was 53.9%. Univariate analysis showed that age ( P=0.033), maximum tumor diameter ( P=0.011), lymph node metastasis ( P=0.002), distant metastasis ( P=0.011), pathological grade ( P=0.001), Mayo staging ( P=0.026) were ralated prognostic factors (all P<0.05). Cox multivariate analysis showed that the pathologically poor differentiation ( HR=1.640, 95% CI 1.112-2.418, P=0.013), and the largest tumor diameter ≥4cm were ( HR=5.000, 95% CI 1.099-22.755, P=0.037). Independent factors affecting patient prognosis. Conclusions:Urachal carcinoma is a malignant bladder tumor with insidient onset. Most of the first diagnosis symptoms are hematuria. When diagnosed, most patients are in the middle and late stages of clinical grading, and the prognosis is poor.Pathological grade and maximum tumor diameter are independent factors that affect the prognosis of patients with urachal carcinoma. The higher was the pathological grade, and the larger was the maximum tumor diameter, the worse was the prognosis.