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Objective:This study aims to analyze the threshold changes in distortion product otoacoustic emissions(DPOAE) and auditory brainstem response(ABR) in adult Otof-/- mice before and after gene therapy, evaluating its effectiveness and exploring methods for assessing hearing recovery post-treatment. Methods:At the age of 4 weeks, adult Otof-/- mice received an inner ear injection of a therapeutic agent containing intein-mediated recombination of the OTOF gene, delivered via dual AAV vectors through the round window membrane(RWM). Immunofluorescence staining assessed the proportion of inner ear hair cells with restored otoferlin expression and the number of synapses.Statistical analysis was performed to compare the DPOAE and ABR thresholds before and after the treatment. Results:AAV-PHP. eB demonstrates high transduction efficiency in inner ear hair cells. The therapeutic regimen corrected hearing loss in adult Otof-/- mice without impacting auditory function in wild-type mice. The changes in DPOAE and ABR thresholds after gene therapy are significantly correlated at 16 kHz. Post-treatment,a slight increase in DPOAE was observeds,followed by a recovery trend at 2 months post-treatment. Conclusion:Gene therapy significantly restored hearing in adult Otof-/- mice, though the surgical delivery may cause transient hearing damage. Precise and gentle surgical techniques are essential to maximize gene therapy's efficacy.
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Ratones , Animales , Emisiones Otoacústicas Espontáneas/fisiología , Audición/fisiología , Oído Interno , Pérdida Auditiva/terapia , Terapia Genética , Umbral Auditivo/fisiología , Potenciales Evocados Auditivos del Tronco Encefálico/fisiología , Proteínas de la MembranaRESUMEN
【Objective】 To investigate the safety and efficacy of flexible ureteroscope (FU) combined with retrograde ureteroscope (URS) in the treatment of lower ureteral calculi with ureteral atresia. 【Methods】 Clinical data of 7 patients treated during Jan.2018 and Jun.2021 were retrospectively analyzed. 【Results】 Ureteral calculi and ureteral atresia were successfully treated, without serious complications. Six to eight weeks after operation, CT examination showed that hydronephrosis was relieved to varying degrees and the ureteral stents were in good position, without residual stones. Six months after operation, there was no significant exacerbation of hydronephrosis and the renal function remained stable. 【Conclusion】 Anterograde FU combined with retrograde URS has high stone-clearance rate, small trauma and high safety in the treatment of ureteral stricture and ureteral calculi.
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Objective: To establish a reliable and sensitive method for evaluating quality of Yiqi Jiangzhi Granules (YQJZG). Methods: Ultra performance liquid chromatography electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) was employed for simultaneous determination of eight marker components. Separation was performed on an AQUITY UPLC® HSS T3 column, the mobile phase consisted of acetonitrile as the organic phase and 0.1% (volume percentage) formic acid as the aqueous. Eight marker components, ginsenoside Rg1 (GRg1), ginsenoside Re (GRe), ginsenoside Rb1 (Gb1), typhaneoside (TEO), isorhamnetin-3-O-neohespeidoside (IN), hesperidin (HPD), aurantio-obtusin-6-O-β-D-glucoside (AG) and curcumin (CCM), were detected by multiple reaction monitoring (MRM) mode. The Chinese Pharmacopoeia (2020 edition) was regarded as the guidance document for this method validation. Results: The method showed good linearity (R
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Objective:To summarize the clinical experience of transurethral columnar balloon dilation of prostate (TUCBDP) in the treatment of patients with benign prostatic hyperplasia(BPH).Methods:A retrospective analysis of 379 BPH clinical data from the Hubei Provincial Hospital of Traditional Chinese Medicine using TUCBDP was performed between June 2015 and June 2018.Their age was (71.3±14.5)years old. The history of disease ranged from 1 month to 36 years. The prostate volume was(47.4±2.1) ml. Preoperative maximum urinary flow rate was (Q max)(9±4) ml/s, postvoid residual urine(PVR) was (123.1±72.4) ml. Their international prostate symptom score (IPSS) was (21±6) points. The quality of life score (QOL)was (5±1) points. The international index erectile function questionnaire (IIEF-5)in 32 patients, who had sex before surgery, was 15±4. We set the time of catheter structure improvement in June 2016 as the boundary, including the early stage (June 2015 to May 2016, 121 cases) and the recent stage (June 2016 to June 2018, 258 patients). In the early stage, the principle of operation is the inner balloon of the catheter to dilate the membrane urethra, and the outer balloon to dilate the urethra of the prostate and the bladder neck. The main surgical steps include the insertion of a dilatation catheter, localization by touching the skin of the scrotum bottom, the inner and outer balloon are filled with water, the first time of drainage and decompression in the inner and outer balloon, the catheter continuous irrigation, drainage and decompression of the inner and outer balloon again, removing the dilatation catheter, and the ordinary urinary catheter was replaced and continuous irrigation. In the recent stage, the principle of surgery is that the inner balloon only served for positioning and fixation. The outer balloon is used to dilate the membrane urethra, prostate urethra, and bladder neck. The inner and outer balloon are drained and decompressed at one time after surgery. The main surgical steps are that the resectoscope was used to examine the bladder and urethra and to guide the dilatation catheter into the bladder. The apex of the prostate touching was used to conform the location. The inner balloon water filling was used for fix the positioning. The inner and outer balloon are filled with water, decompressed and pulled out for urination test, the gland expansion is observed under the resectoscope, and ordinary urinary catheter is replaced for continuous flushing. We observed the changes in Q max, PVR, IPSS, and QOL at 1, 3, 6, 12, and 24 months after the operation. the complications differences in two-stage patien, including the International Incontinence Advisory Committee Urinary Incontinence Questionnaire (ICI-Q-SF) score; those who had sex before surgery were recorded changes in the IIEF-5 score, was compared. Results:There were no deaths during and after operation in this study. The operation time was (18.5±6.7) min. The number of follow-up cases at 1, 3, 6, 12, and 24 months after operation were 326, 253, 201, 194, and 181, respectively. The Q max at 1, 3, 6, 12, and 24 months after operation were (17±9)ml/s, (15±2)ml/s, (12±4)ml/s, (13±6)ml/s and (13±4)ml/s, respectively. The PVR were (17.4± 11.6) ml, (20.6±9.8)ml, (25.4±13.1)ml, (31.5±11.5)ml, and (29.1±12.4)ml, respectively. The IPSS were(7±5) points, (4±4) points, (4±4) points, (6±5) points, (4±4) points, respectively. The QOL were (2±1) points, (2±1) points, (2±1) points, (2±1) points, and (2±1), respectively. All those results that were significantly different from those before surgery ( P<0.05). There were 32 patients who had sex before the operation. The postoperative IIEF-5 score was (17± 6), which was not significantly different from that before the operation ( P>0.05). Two patients had transient retrograde ejaculation, which relieved spontaneously within the 6 month. 4 cases with pseudourinary incontinence in the recent stage (1.5%) were not statistically different from 6 cases (4.9%) in the early stage ( P>0.05). one case(0.4%) of major bleeding in the recent stage was statistically different from 6 cases (4.9%) in the early stage ( P<0.05). 2 cases (0.7%) of patients with acute urinary retention in the recent stage were significantly different from 15 cases (12.4%) in the early stage ( P<0.05). Conclusions:TUCBDP has a positive overall effect and high safety. The major complications of surgery in the recent stage, except for pseudo-urinary incontinence, are significantly lower than that in the early stage, which may be related to the improvement of the catheter structure and the accumulation of clinical experience.
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AIM: To investigate the inhibitory effect of vector-based RNA interference(RNAi) on the expression of melanoma associated antigen A3(MAGEA3) protein in hepatocellular carcinoma cells and on apotposis of hepatocellular carcinoma cells.METHODS: A vector for transcribing specific small hairpin RNA(shRNA) targeting MAGEA3 gene was constructed,introduced into hepatocellular carcinoma MEL-ED1 cells by Lipofectamine 2000.The MAGEA3 protein and mRNA expression levels of MEL-ED1 cells were detected by Western blotting and RT-PCR, respectively.The cell apoptosis was studied by DNA fragmentation,electron microscopy,TUNEL assay,and annexin V/PI staining.RESULTS: The vector of RNA interference was successfully constructed and MAGEA3 expression was descreased significantly in MEL-ED1 cells.After the shRNA expression vector was transfected into the MEL-ED1 cells,the expression of MAGEA3 gene was inhibited significantly(by 90%).DNA fragmentation,electron microscopy and TUNEL assay showed classic apoptosis characters in the MEL-ED1 cells transfected with pSilencer-MAGEA3 plasmid with an apoptosis rate of 21.41% ?1.98%,significantly higher than those in the negative control group transfected with pSilencer-neo and in the non-transfected group(both P