OBJECTIVE:To develop a method for simultaneous determination of chlorogenic acid,geniposide,gentiopicro-side,ferulic acid,baicalin and ammonium glycyrrhizinate in Yindan pinggan capsule. METHODS:HPLC method was adopted. The separation was performed on Wonda SilTM-C18 column with mobile phase consisting of acetonitrile-0.4% phosphoric acid solu-tion(gradient elution)at the flow rate of 0.8 mL/min. The detection wavelength were set at 325 nm(chlorogenic acid,ferulic ac-id),250 nm (geniposide,ammonium glycyrrhizinate) and 275 nm (gentiopicroside,baicalin). The column temperature was 30 ℃. RESULTS:The linear ranges were 0.087-3.480 μg for chlorogenic acid(r=0.9998),0.201-8.040 μg for geniposide(r=0.9997),0.200-8.000 μg for gentiopicroside(r=0.9995),0.016-0.640 μg for ferulic acid(r=0.9999),0.105-4.200 μg for ba-icalin (r=0.9999) and 0.028-1.120 μg for ammonium glycyrrhizinate (r=0.9995),respectively. The limits of quantitation were 1.31,0.75,1.14,1.25,0.94,0.98 ng,and the limits of detection were 0.87,0.67,0.96,0.93,0.60,0.88 ng,respectively. RSDs of precision,stability and reproducibility tests were lower than 2.0%;recoveries were 99.9%-101.9%(RSD=0.7%,n=6), 98.7%-100.9%(RSD=0.9%,n=6),98.1%-101.5%(RSD=1.4%,n=6),98.5%-101.3%(RSD=1.3%,n=6),98.5%-101.7%(RSD=1.2%,n=6),98.2%-101.4%(RSD=1.2%,n=6),respectively. CONCLUSIONS:The method is simple and accurate , can be used for simultaneous determination of chlorogenic acid,geniposide,gentiopicroside,ferulic acid,baicalin and ammonium glycyrrhizinate in Yindan pinggan capsule.

BACKGROUND:The process of oxidative stress that impacts the curative effect exists in the region which accepts cel transplantation. However, there are few reports about the effects of oxidative stress on human dental pulp stem cels and relevant mechanism.
OBJECTIVE:To understand the effect of hydrogen peroxide on the senescence of human dental pulp stem cels.
METHODS:Human dental pulp stem cels were isolated and cultured in PBS, 100 and 200 μmol/L hydrogen peroxide for 2 hours, respectively. Cel morphology was observed under inverted microscope, degree of cel senescence monitored by β-galactosidase staining, cel proliferation ability detected by BrdU kit and cel counting method, cytoskeleton of dental pulp stem cels and expression of sirt1 tested using immunofluorescence method, and expression of sirt1 and p16 proteins measured by western blot assay.
RESULTS AND CONCLUSION:Dental pulp stem cels exhibited a fibroblast-like morphology with spindle-shaped appearance. After stimulated by hydrogen peroxide, the cel volume was enlarged, theβ-galactosidase staining deepened and the proliferation of dental pulp stem cels reduced. The enhancement of senescence of dental pulp stem cels was accompanied with the increasing concentration of hydrogen peroxide, and in this process, the expression of p16 was raised while the expression of sirt1 was decreased. In conclusion, the senescence of human dental pulp stem cels can be promoted by the stimulation of hydrogen peroxide, and sirt1 and p16 are involved in this process. Our findings may provide a theoretical and experimental foundation for autologous transplantation of dental pulp stem cels.

BACKGROUND:During the healing of fractures, removal of sciatic nerve can result in insufficient mechanical rigidity of newborn woven bone. However, there are less reports concerning the denervation effects during distraction osteogenesis. OBJECTIVE:To observe the effect of removal of the sciatic nerve on bone regeneration and the expression of Runt-related transcription factor 2 (Runx2) protein during distraction osteogenesis in a rabbit model. METHODS:Twenty-four adult male New Zealand rabbits were selected and underwent left tibial osteodistraction to construct animal models of distraction osteogenesis. Before distraction, the animals were randomly divided into group R (resecting the left sciatic nerve) and group I (intact left sciatic nerve). Six weeks after completion of distraction, the animals were kil ed and the lengthened tibias were harvested for radiography, three-dimensional CT reconstruction, histological evaluation, connectivity density (Conn.D) evaluation. RESULTS AND CONCLUSION:New regenerated bone was present and Runx2 protein was expressed in the distraction gaps of al animals at the end of the study, as revealed by radiography, three-dimensional CT reconstruction, and histological observation. However, less new bone formation and a lower degree of mineralization and expression of Runx2 protein were observed in group R compared with group I. The results suggest that the denervation appears to have an inhibitory effect on bone formation and the expression of Runx2 protein during distraction osteogenesis.

Chronic tympanic membrane perforation in an animal model has been widely used. The ideal model is a lasting and stable perforation without infection. For half a century, several physical modalities have been evolved, such as thermal injury, infolding technique, tympanostomy tube, re-myringotomy, and laser myringotomy. Chemical methods using chemical substance inhibiting growth and repair of cells as well as using gene defect animals are also applied. We have found the success rate is lower when using only one method and it would be better to combine physical and chemical approaches. In this article, we review the research advances in the establishment of chronic tympanic membrane perforation animal model.

For otitis media patients with severe mucosal lesions, adhesion is the most common poor prognosis of tympanoplasty. We generally consider tympanic cavity adhesion is due to eustachian tube dysfunction and poor middle ear ventilation. The mechanism of adhesion is unclear so far, which we thought is mainly associated with wounded surface and activation of the fibrinolytic system. To solve the problem of adhesion, several materials have been used in the middle ear surgery, such as plastic sheet, silicone sheet and absorbable material. And there are some benefits. The most widely used is silicone sheet, and there are some researches focused on its thickness, shape and components to achieve better anti-adhesion effect. In this article, we will do a review for the research advances of anti-adhesion materials in tympanoplasty.

Objective:To investigate the technical points and clinical application of combined infratemporal fossa approaches (CIFA) by analyzing the clinical results with CIFA for lesions involved skull base.Methods:A retrospective study was performed on 11 patients underwent CIFA for skull base lesions dissection from December 2014 to January 2019 in the department of otolaryngology, Peking Union Medical College Hospital. There were 6 male and 5 female patients, with age range of 16-72 years old and median age of 53 years old. Five patients underwent CIFA Type B and D, and the other 6 underwent CIFA A and B. All patients were followed up regularly by CT and MRI to observe possible recurrence.Results:Among the 5 patients with CIFA Type B and D, 3 were giant cell tumor and 2 were giant cell reparative granuloma, and median maximum cross-section size was 42 mm×46 mm (range from 37 mm×18 mm to 56 mm×53 mm). Among the 6 patients with CIFA Type A and B, 4 were paraganglioma of head and neck, 1 was schwannoma of skull base, 1 was petrous cholesteatoma, and median maximum cross-section size was 43 mm×36 mm (range from 24 mm×22 mm to 63 mm×35 mm). Nine patients underwent complete resection of the tumor in the first stage. In 2 patients, the extracranial parts were removed in the first stage, and the intracranial part was removed in the second stage. Tympanum and ossicular reconstruction were done in one of the CIFA Type B and group D, and 1 year′s postoperative hearing was mild conductive hearing loss. There was no cerebrospinal fluid leakage of all patients. All the 5 patients with normal facial nerve function before surgery recovered to H-B grade Ⅰ to Ⅱ within 3 months after surgery. Among the 4 patients whose preoperative facial nerve function were grade Ⅱ, 2 recovered to grade I after surgery and the other 2 were still grade Ⅱ. For the patient whose preoperative facial nerve function was grade Ⅴ, his postoperative recovery was grade Ⅲ. There was 1 patient whose pre-operative FN function was H-B grade Ⅲ, and the post-operative FN function was grade Ⅵ due to FN resection. Except for 2 cases with cochlear involved before surgery, cochleae of the other 9 cases were preserved. The follow-up time was 14 to 58 months. No recurrence was observed in all patients.Conclusions:The CIFA can safely and completely remove the extensive lesions that invade the skull base, and the facial nerve function can be well protected and recovered intro-and post-operation. Appropriate use of combined IFA can not only achieve good exposure and complete resection of lesions, but also create conditions for functional reconstruction.

Objective:To analyze the efficacy of unilateral arytenoid chondroplasty combined with minimally invasive mucosal flap plasty in the management bilateral vocal cord paralysis.Methods:A total of 66 patients with bilateral vocal cord paralysis hospitalized in the First Hospital of Sun Yat-sen University from January 2018 to December 2023 were retrospectively analyzed, among whom there were 8 males and 58 females, with ages ranging from 35 to 86 years old(mean age (57.8±11.6) years). All patients underwent suspension laryngoscopic CO 2 laser unilateral arytenoid chondroplasty and mucosal flap under general anesthesia. Postoperative follow-up period extended from 6 months to 6 years, with a median duration of 28 months. The study compared the degree of dyspnea and voice quality (subjective and objective evaluation) of the patients pre- and post-operatively, and analyzed the clinical differences between patients with and without preoperative or intraoperative tracheotomy, the extubation rate of tracheotomized patients, the recurrence rate, and the complication rate. Continuous variables conforming to normal distribution were tested by t-test and categorical variables by χ2 test,the Wilcoxon rank-sum test was used to analyze the improvement in dyspnea before and after surgery. Results:Compared with the preoperative period, 59 patients showed improvement in postoperative dyspnea ( U=161.5, P<0.01); there was no significant difference in voice disorder index 10, subjective auditory-perceptual assessment, and maximal vocalization time ( P>0.05), all of which remained within the relative normal range. Tracheotomy was performed in 32 out of 66 patients, with predominantly degree Ⅲ- dyspnea (46.9%, 15/32), including 26 patients with preoperative tracheotomy and 6 patients with intraoperative tracheotomy. Among the 34 patients who did not undergo tracheotomy, the majority presented with degree Ⅱ-dyspnea (82.4%, 28/34). All patients achieved successful extubation following surgery, with a mean median time to extubation of 1 month. Conclusions:The combination of unilateral arytenoid chondroplasty and minimally invasive mucosal flap plasty represents a refined and effective therapeutic approach for bilateral vocal cord paralysis. This minimally invasive technique has the potential to reduce the rate of tracheotomy.

【Objective】 To investigate the practice and benefit of national standardized management of type 2 diabetes in Yulin City. 【Methods】 We recruited the adult type 2 diabetes patients who sought medical help at our hospital from May 2020 to October 2022 as subjects. We collected their basic information (sex and age); measured height, weight, waist and hip circumference, and blood pressure; calculated body mass index (BMI); and detected blood glucose, c-peptide, HbA1c, biomarkers, urinary microalbumin, sensory nerve conduction velocity of lower limbs, ABI, and subcutaneous and visceral fat at the time of MMC recruited and the end of six months. T test and Mann-Whitney U rank sum test were used for measurement data and χ² test or Fisher’s exact probability method for counting data to analyze the data. 【Results】 After 6 months, the levels of fasting blood glucose, postprandial blood glucose, HbA1c, and visceral and subcutaneous fat in all the patients decreased, but the level of fasting c-peptide increased compared with the baseline (all P<0.05). Secondly, compared with the baseline, the control rate of HbA1c (35.21% vs. 13.71% ) and the comprehensive control rate (13.97% vs. 7.26% ) were both significantly increased at six months (P<0.05). Thirdly, after 6 months, the levels of fasting blood glucose, postprandial blood glucose, HbA1c, TG, TC, and UA were decreased more, while the fasting c-peptide and postprandial c-peptide were increased more in the patients of the HbA1c standard group (HbA1c<7% ) than those of the non-standard group. 【Conclusion】 The multiple benefits of blood glucose, blood lipid, uric acid and islet function can be achieved by taking type 2 diabetes patients into MMC. Meanwhile, the rates of HbA1c control and comprehensively reaching the standard are significantly increased. Therefore, MMC can explore a new way for the management of type 2 diabetic patients in this area.

Extracellular vesicles (EVs) have recently received much attention about the application of drug carriers due to their desirable properties such as nano-size, biocompatibility, and high stability. Herein, we demonstrate orange-derived extracellular vesicles (OEV) nanodrugs (DN@OEV) by modifying cRGD-targeted doxorubicin (DOX) nanoparticles (DN) onto the surface of OEV, enabling significantly enhancing tumor accumulation and penetration, thereby efficiently inhibiting the growth of ovarian cancer. The obtained DN@OEV enabled to inducement of greater transcytosis capability in ovarian cancer cells, which presented the average above 10-fold transcytosis effect compared with individual DN. It was found that DN@OEV could trigger receptor-mediated endocytosis to promote early endosome/recycling endosomes pathway for exocytosis and simultaneously reduce degradation in the early endosomes-late endosomes-lysosome pathway, thereby inducing the enhanced transcytosis. In particular, the zombie mouse model bearing orthotopic ovarian cancer further validated DN@OEV presented high accumulation and penetration in tumor tissue by the transcytosis process. Our study indicated the strategy in enhancing transcytosis has significant implications for improving the therapeutic efficacy of the drug delivery system.