Objective:To assess the prognostic value of methylation of interferon regulatory factor 6 ( IRF6) gene promoter in patients diagnosed with Kidney renal clear cell carcinoma (KIRC). Methods:The primary lesions of fifty KIRC patients who were diagnosed at the First Affiliated Hospital of Nanjing Medical University from January 2016 to January 2020 were collected. The expression of IRF6 protein was determined with an immunohistochemical method. The correlation between the level of IRF6 expression and survival and/or metastasis status was analyzed. The mRNA and protein levels of the IRF6 in KIRC and normal renal tissues were compared by using bioinformatic tools. The difference in the methylation rate of the IRF6 gene promoter between tumor and adjacent tissues was analyzed by searching the online databases. Statistical analysis was carried out for the methylation status of the IRF6 gene promoter region to select those negatively correlated with the overall survival (OS) among the patients. In vitro experiments were conducted with cell lines to verify the correlation between the status of promoter methylation and transcription level of the IRF6 gene. Results:The mRNA and protein levels of the IRF6 gene in KIRC tissues were significantly lower than those of the normal controls, and this was more prominent in patients who had died or developed metastasis. The extent of IRF6 gene promoter methylation in the KIRC tissues was much higher compared with that of the adjacent normal renal tissues. There was a significant negative correlation between the methylation of the IRF6 gene promoter and mRNA level of the IRF6 ( R= -0.52). The higher methylation degree in the IRF6 gene promoter regions cg12034118 and cg16030177, the shorter the OS and worse prognosis in the patients. Only twenty CpG sites in cg12034118 were confirmed to be highly methylated in KIRC cell lines. The transcription level of the IRF6 gene was upregulated in a time- and dose-dependent manner after the treatment with demethylation reagent 5-azadeoxycytidine. Conclusion:The methylation of IRF6 gene promoter in the renal tissues of KIRC patients is closely correlated with the OS. Cg12034118 may provide a promising biomarker for laboratory detection, and its high methylation rate has certain reference value for the prognosis.

DNA-encoded chemical library (DEL) links the power of amplifiable genetics and the non-self-replicating chemical phenotypes, generating a diverse chemical world. In analogy with the biological world, the DEL world can evolve by using a chemical central dogma, wherein DNA replicates using the PCR reactions to amplify the genetic codes, DNA sequencing transcripts the genetic information, and DNA-compatible synthesis translates into chemical phenotypes. Importantly, DNA-compatible synthesis is the key to expanding the DEL chemical space. Besides, the evolution-driven selection system pushes the chemicals to evolve under the selective pressure, i.e., desired selection strategies. In this perspective, we summarized recent advances in expanding DEL synthetic toolbox and panning strategies, which will shed light on the drug discovery harnessing in vitro evolution of chemicals via DEL.

Objective:To investigate the safety and efficacy of ultrasound-guided percutaneous radiofrequency ablation in the treatment of plasma cell mastitis.Methods:A retrospective analysis was performed for the clinical data of 10 patients with plasma cell mastitis treated with ultrasound-guided percutaneous radiofrequency ablation in the First Affiliated Hospital of Zhengzhou University from January 2021 to December 2022. The postoperative complications, symptoms and signs of the patients, ultrasound imaging manifestations, treatment efficacy at 3 months after surgery, the reduction rate of ablation lesions at 1 month, 3 months, 6 months, and 12 months after surgery, the recurrence status within 1 year after surgery, and the satisfaction with the treatment effect were observed.Results:The reduction rates of ablation lesions at 1 month, 3 months, 6 months, and 12 months were 32.31%-51.00%[(42.78±4.48)%], 70.66%-86.68%[(70.45±12.43)%], 72.31%-100%[(86.91±7.45)%], 89.13%-100%[(96.07±7.45)%], respecctively. The symptoms and signs of 10 patients improved significantly 3 months after surgery, and there was no recurrence of the disease during the follow-up period, and the treatment effect was significant and satisfactory.Conclusions:Ultrasound-guided percutaneous radiofrequency ablation for the treatment of plasma cell mastitis is a minimally invasive, effective and safe treatment.

Objective:To compare the therapeutic effects of ultrasound-guided radiofrequency ablation (RFA) and surgical resection on salivary gland mixed tumor with diameter ≤3 cm, and to analyze the feasibility and clinical value of RFA in the treatment of salivary gland mixed tumor.Methods:A retrospective study was conducted on 43 patients with salivary gland mixed tumor whose diameter ≤3 cm were treated in the First Affiliated Hospital of Zhengzhou University from March 2020 to March 2022.According to the treatment method, they were divided into an ablation group of 20 cases and a resection group of 23 cases. The complete ablation rate of the ablation group was evaluated, the postoperative absorption of ablation foci in the ablation group was recorded, and the perioperative indexes and the incidence of postoperative complications of the two groups were compared. The long-term prognosis of the two groups was followed up, and the incidence of recurrence, malignant metastasis and appearance recovery were evaluated.Results:In the ablation group, the complete ablation rate was 100.00% (20/20). The lesion volume reduction rates at 1, 3, 6 and 12 months after operation were -(39.60±16.18)%, (8.61±8.10)%, (31.87±14.68)% and (58.64±14.82)%, respectively. The operation time, recovery feeding time, postoperative hospital stay and intraoperative bleeding in the ablation group were less than those in the resection group, and the differences were statistically significant (all P<0.001). Postoperative complications occurred in 2 cases in the ablation group and 9 cases in the resection group. There was significant difference in the total incidence of postoperative complications between the two groups(χ 2=4.768, P=0.029). There was 1 case of recurrence in the ablation group, and no facial depression or protuberance occurred 1 year after operation. There was no recurrence in the resection group, one year after operation, the incidence of facial depression and facial protuberance was 13.04%(3/23) and 0, respectively.There was no malignant metastasis in both groups. Conclusions:Ultrasound-guided RFA is safe and effective in the treatment of mixed tumors of salivary gland with diameter ≤3 cm, and the therapeutic effect is not significantly different from that of surgery. With minimal trauma and good cosmetic effect, it has a great application prospect.

Combination of passive targeting with active targeting is a promising approach to improve the therapeutic efficacy of nanotherapy. However, most reported polymeric systems have sizes above 100 nm, which limits effective extravasation into tumors that are poorly vascularized and have dense stroma. This will, in turn, limit the overall effectiveness of the subsequent uptake by tumor cells via active targeting. In this study, we combined the passive targeting via ultra-small-sized gemcitabine (GEM)-based nanoparticles (NPs) with the active targeting provided by folic acid (FA) conjugation for enhanced dual targeted delivery to tumor cells and tumor-associated macrophages (TAMs). We developed an FA-modified prodrug carrier based on GEM (PGEM) to load doxorubicin (DOX), for co-delivery of GEM and DOX to tumors. The co-delivery system showed small particle size of ∼10 nm in diameter. The ligand-free and FA-targeted micelles showed comparable drug loading efficiency and a sustained DOX release profile. The FA-conjugated micelles effectively increased DOX uptake in cultured KB cancer cells that express a high level of folate receptor (FR), but no obvious increase was observed in 4T1.2 breast cancer cells that have a low-level expression of FR. Interestingly, in vivo, systemic delivery of FA-PGEM/DOX led to enhanced accumulation of the NPs in tumor and drastic reduction of tumor growth in a murine 4T1.2 breast cancer model. Mechanistic study showed that 4T1.2 tumor grown in mice expressed a significantly higher level of FOLR2, which was selectively expressed on TAMs. Thus, targeting of TAM may also contribute to the improved in vivo targeted delivery and therapeutic efficacy.

Objective:To investigate the safety and efficacy of ultrasound-guided percutaneous radiofrequency ablation in the treatment of nonmetastatic pheochromocytoma.Methods:A retrospective analysis was performed on 7 patients with nonmetastatic pheochromocytoma admitted to the First Affiliated Hospital of Zhengzhou University from January 2018 to June 2020, all of whom underwent ultrasound-guided percutaneous radiofrequency ablation. The changes of postoperative blood pressure, improvement of symptoms, intraoperative and postoperative complications were observed.Plasma free methoxypinephrine (MN) and normetanephrine (NMN) levels were recorded before and 2 weeks after operation. The reduction rate of ablation lesion volume at 1, 3 and 6 months after operation was calculated.Results:Postoperative blood pressure of all 7 patients was reduced to the normal range within 3 days, and symptoms such as headache were significantly relieved immediately after operation.No serious complications occurred during or after operation. Plasma free MN and NMN levels decreased to normal levels 2 weeks after operation. The mean reduction rates of the ablation lesions at 1, 3 and 6 months after operation were (46.61±13.42)%, (67.21±10.54)% and (85.73±4.15)%, respectively. Postoperative follow-up of 12-30 months showed that the blood pressure, plasma free MN and NMN levels of the patients were all in the normal range, and no symptoms such as headache and palpitation occurred again. All the tumors were completely ablated, and no recurrence was observed.Conclusions:Ultrasound-guided percutaneous radiofrequency ablation in the treatment of nonmetastatic pheochromocytoma is minimally invasive, safe and effective, and can retain adrenal cortex function, which is worthy of clinical promotion.

Objective:To investigate the clinical efficacy of ultrasound-guided percutaneous radiofrequency ablation in the treatment of ectopic pregnancy.Methods:A retrospective analysis was performed on 8 patients with ectopic pregnancy admitted to the First Affiliated Hospital of Zhengzhou University from January 2018 to January 2020, all of whom underwent ultrasound-guided percutaneous radiofrequency ablation. Contrast ultrasonography was performed immediately after the operation to observe the changes of pregnancy sac. The length of hospital stay, mass absorption time and menstrual recovery time of the patients were recorded. Postoperative complications and serum human chorionic gonadotropin (HCG) negative conversion time were observed. At 3 months after the operation, hysterosalpingography was performed 3-7 days after the menstruation to observe the patency of the fallopian tubes, all patients were followed up for 1 year to observe the postoperative intrauterine pregnancy and re-ectopic pregnancy.Results:All the 8 patients were successfully treated with radiofrequency ablation, and postoperative ultrasonography showed that the mass remained unenhanced. The length of hospital stay was (3.0±0.8)d. The packet absorption time was (32.4±14.3)d. The recovery time of menstruation was (39.6±2.7)d. There were no serious complications.Blood HCG levels decreased rapidly within a week, and all returned to normal level of non-pregnancy 2-3 weeks after the operation. Three months after the operation, the salpingogram showed that the fallopian tubes were recanalized in 3 patients. After a follow-up of 1 year, 1 patient had a natural intrauterine pregnancy, and there was no case of recurrent ectopic pregnancy.Conclusions:Ultrasound-guided percutaneous radiofrequency ablation is effective in the treatment of ectopic pregnancy with minimal trauma and rapid recovery, and can retain the reproductive ability of patients to some extent, which is worth popularizing in clinical application.

Occupational Exposure ; prevention & control ; Risk Assessment ; methods

Objective To observe the prokinetic effect of domperidone on esophagus and lower esophageal sphincter (LES), and compare its effect with that of mosapride and cisapride. Methods In vivo experiments: forty rats were divided into control, domperidone, mosapride, and eisapride groups. Strain gauges were planted in proximal esophagus, distal esophagus and LES to record the activities of esophagus and LES in conscious rat. In vitro experiments: in the thermostatic muscle bath, the prokinetic effect of domperidone, mosapride, and cisapride on the contractility of rat muscle strips from esophagus body and LES were recorded by tone-transducers. Results In vivo experiments, ① In the interdigestive period of resting conscious rats, only mild contraction activities were recoded in esophagus bodies. In LES, typical interdigestive migrating motor complex (MMC) with phase Ⅰ,Ⅱ,Ⅲ,and Ⅳ was recorded. The contraction amplitude of LES was much greater than esophagus body. ② Domperidone significantly enhanced the contraction of esophagus body and LES. The mean contraction amplitude of proximal esophagus, distal esophagus, and LES increased by 63.24%±7.17%, 75.54%±5.27%, and 85.81%±6.02%, respectively, compared with controls. The prokinetic effect showed a dose-effect relation. Mosapride at the same dosage increased the mean contraction amplitude of proximal esophagus, distal esophagus, and LES by 29. 71%±4.15%, 40.15%±3.30%, and 35.24%±5.36%, respectively, compared with controls. The prokinetic effects of mosapride on esophagus and LES were much less than domperidone. Cisapride at the same dosage increased the mean contraction amplitude of proximal esophagus, distal esophagus, and LES by 59.84%±6.55%, 70.11%±5.62%, and 75.13%± 5.10%, respectively, compared with controls. The prokinetic effects of cisapride were similar as domperidone. In vitro experiments. ① Domperidone perfusion could significantly increase the contraction of esophagus body and LES muscle strips by 87.74%±7.65% and 92.44±7.17%, respectively, compared with Krebs-Ringer (KR) solution perfuslon. Mosapride at the same dosage increased the mean contraction amplitude by 35.42%±5.02% and 31.12%±4.32%, respectively, compared with KR controls. The prokinetic effects of mosapride were much less than domperidone. Cisapride of the same dosage showed a similar prokinetic effect as domperidone. ② Atropine and tetrodotoxin could block the prokinetic effects of domperidone on esophagus and LES. Conclusions Domperidone can significantly enhance the esophagus body contraction and LES motility. The effects of domperidone are similar as cisapride and much greater than mosapride. The prokinetic effects of domperidone on esophagus and LES are not only through well-known dopaminergic receptor blockade, but also through the cholinergic nerves of the enteric nervous system.

Objective To investigate the diagnostic value of JAK2V617F mutation detection in the diagnosis of polycythemia vera,and screen a more simple method for clinic laboratory to detect the mutation of JAK2V617F.Method DNA was extracted by standard procedures after isolating total leukocytes from peripheral blood mononuclear cells by density gradient centrifugation over Histopaque 1077.DNA samples were amplified,and single-stranded biotinylated PCR products were prepared for sequencing.At the same time,in order to testify the reliability of the allele-specific PCR,two forward primer and one common reverse primers were applied for identifying the mutation.Result In 32 of 38 patients with polycythemia vera,JAK2V617F mutation was determined by conventional DNA sequencing.35 JAK2V617F mutations were detected by the allele-specific PCR.and these mutations were confirmed by DNA sequencing.None mutation was found in secondary polycythemia and normal control.Conclusion JAK2V617F function mutation occuis in nearly all patients with PV,and JAK2V617F mutation should be a molecular marker in the diagnosis of polycythemia vera.Allele-specific PCR is a very sensitive and specific method for detecting JAK2V617F mutation.