ABSTRACT
Objective:To explore the feasibility and safety of the clinical application of the diagnosis and treatment mode combining rapid frozen pathological examination of prostate biopsy tissue with radical prostatectomy.Methods:Suspected prostate cancer patients with PSA>10 ng/ml and PI-RADS score ≥4 in, Northern Jiangsu People's Hospital from April to September 2021 were collected. The included patients underwent mpMRI/TRUS image fusion-guided transperineal prostate targeted biopsy with 16G biopsy needle, 2-3 needles for biopsy, and rapid frozen pathological examination. Robot-assisted laparoscopic radical prostatectomy (RALP) was performed immediately for patients with prostate cancer with rapid freezing pathology. For undiagnosed prostate cancer, 18G biopsy needle for prostate targeted + systematic biopsy were used, 18-22 needles for systematic biopsy, and routine pathological examination. The baseline data, frozen pathological results, perioperative conditions, pathological results and follow-up data of all patients were collected.Results:Eleven patients were included in the study, the mean age of the patients was 69.9(66-73) years, the mean BMI was 22.8(19-26) kg/m 2, the mean PSA was 23.2(14.25-32.00), the mean prostate volume was 45(32-52) ml, mean PSAD 0.54(0.33-0.75). PI-RADS score was 4 in 3 cases and 5 in 8 cases; digital rectal examination was positive in 5 cases. All 11 cases underwent rapid freezing and the pathological results showed that: 9 cases were prostate adenocarcinoma, and RALP was performed immediately. The operation time was 111.5(96-126) min, the intraoperative blood loss was 78.9(55-105) ml, and the postoperative extubation time was 4.3(3.5-5.0) days, postoperative hospital stay 5.8(5.0-6.5) days. Postoperative pathology showed that Gleason score 3+ 4=7 in 1 case, 4+ 3=7 in 3 cases, 8 points in 4 cases, and 10 points in 1 case; 3 cases had positive resection margins, and 1 case had seminal vesicle invasion, the average number of dissected lymph nodes was 10.9 (8.5-14.0), and there was no tumor metastasis. Pathological T staging included 2 cases of T 2b stage, 5 cases of T 2c stage, 1 case of T 3a stage, and 1 case of T 3b stage. Two patients were undiagnosed by rapid freezing pathology, of which one was prostate adenocarcinoma with a Gleason score of 4+ 3=7, and then received RALP; the other one was prostate inflammation. 11 patients were followed up; the postoperative follow-up time was 3-7 months, with an average of 5.2 months. Among the 10 patients who underwent RALP, 8 patients recovered urinary continence 2 weeks after surgery, and all recovered within 2 months after surgery. Three patients with positive surgical margins were given regular androgen deprivation therapy in the second week after surgery. PSA did not drop below 0.1 ng/ml in patients with positive margins and seminal vesicle invasion 3 months after surgery. No complications of Clavien grade Ⅰ or higher occurred after operation and during follow-up. Conclusions:For patients with high suspicion of prostate cancer, rapid frozen pathological examination of prostate biopsy tissue is performed. RALP is performed immediately for patients with prostate cancer. The results show that this diagnosis and treatment model could be safe and feasible.
ABSTRACT
OBJECTIVE: To compare our in-house method of embryo freezing with Cryotop vitrification in terms of immediate survival, subsequent cleavage and blastocyst formation, and cell numbers in blastocysts. METHODS: Two-cell mouse embryos were randomly allocated into three groups: a non-frozen control group (group 1, n=300), a group that underwent Cryotop vitrification (group 2, n=300), and a group that underwent our in-house freezing method (group 3, n=300). RESULTS: There were no significant differences between groups 2 and 3 in the immediate survival rate (96.3% vs. 98.6%, respectively; p=0.085), the further cleavage rate (91.7% vs. 95.0%, respectively; p=0.099), or the blastocyst formation rate (80.7% vs. 78.6%, respectively; p=0.437). The cell numbers in the blastocysts from groups 1, 2, and 3 were comparable (88.99±10.44, 88.29±14.79, and 86.42±15.23, respectively; p=0.228). However, the percentage of good-quality blastocysts in the Cryotop vitrification group was significantly higher than in the group in which our in-house method was performed, but was lower than in the control group (58.0%, 37.0%, and 82.7%, respectively; p < 0.001). CONCLUSION: At present, our method is inferior to the commercial Cryotop vitrification system. However, with further improvements, it has the potential to be useful in routine practice, as it is easier to perform than the current vitrification system.
Subject(s)
Animals , Mice , Blastocyst , Cell Count , Embryonic Structures , Freezing , Methods , Survival Rate , VitrificationABSTRACT
El incremento del número de pacientes que desean mantener su fertilidad, ya sea por motivos oncológicos o de fertilidad, como son los pacientes con enfermedades infecciosas virales trasmitidas por vía sexual, o que se someten en forma voluntaria a la esterilización quirúrgica, requieren de métodos de congelación que preserven en forma adecuada la función de los espermatozoides. En el área de la criobiología, la utilización de técnicas de congelación ultrarrápida ha permitido preservar en forma exitosa ovocitos, embriones y tejido ovárico. Este método se ha incorporado recientemente para preservar el gameto masculino. El presente estudio evalúa el efecto de la congelación ultrarrápida (vitrificación) sobre la función espermática de 10 donantes normozoospérmicos. Los espermatozoides se seleccionaron por Swim-up y la solución espermática se dividió en dos subfracciones. Una fracción se vitrificó sumergiéndola directamente en nitrógeno líquido mientras que la segunda se utilizó como control. En ambas fracciones se determinaron viabilidad, movilidad, potencial de membrana mitocondrial (YMMit), integridad del ADN, reacción de acrosoma espontánea e inducida, y superóxido intracelular (O2.-). Se observó que la vitrificación preserva una adecuada función celular en un alto número de espermatozoides, siendo además un método simple, rápido y de menor costo, ya que no necesita equipo de congelación. No obstante, existe una significativa activación de la producción de especies reactivas de oxígeno, que conlleva a una prematura capacitación espermática, evento que es necesario de modular, especialmente si se utilizan estas células en técnicas de inseminación intrauterina. Futuros estudios con adición de antioxidantes a los medios de congelación parecen necesarios para optimizar esta técnica.
The number of patients who wish to maintain their fertility is ever increasing. This group of patients includes cancer patients, those with fertility problems or viral infectious diseases acquired through sexual contact and others submitting to voluntary surgical sterilization; all of the above requiring freezing methods to adequately preserve sperm function. In the field of cryobiology the use of ultra-rapid freezing techniques has successfully preserved oocytes, embryos and ovarian tissue. This method has recently been incorporated in preserving male gametes. This study evaluates the effect of ultra-rapid freezing (vitrification) on sperm function of 10 normozoospermic donors. The sperm were selected by swim-up technique and the solution divided into two fractions. One fraction is vitrified by dipping directly into liquid nitrogen and the second fraction is used as control. In both fractions, viability, motility, mitochondrial membrane potential (YMMit) DNA integrity, spontaneous and induced acrosome reaction and intracellular superoxide (O2.-) were determined. It was noted that vitrification preserves cell function in a great number of spermatozoon, and is also simple, rapid and cost effective as this method does not require freezing equipment. There is however, significant activation of the production of reactive oxygen species, which leads to premature sperm capacitation, an event necessary to modulate particularly when using these cells in intrauterine insemination techniques. Future studies with addition of antioxidants to freezing media are necessary to further improve this technique.
Subject(s)
Adult , Cryopreservation/methods , Spermatozoa/cytology , Spermatozoa/physiology , Spermatozoa/ultrastructure , Semen Preservation/methods , Sperm Banks/methodsABSTRACT
The purpose of this study was to evaluate the viability of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure by means of MTT assay, WST-1 assay. 12 teeth of Sprague-Dawley white female rats of 4 week-old were used for each group. Both side of the first and second maxillary molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group 1 (Immediate extraction), group 2 (Slow freezing under pressure of 3 MPa), group 3 (Slow freezing under pressure of 2 MPa), group 4 (Slow freezing under no additional pressure), group 5 (Rapid freezing in liquid nitrogen under pressure of 2 MPa), group 6 (Rapid freezing in liquid nitrogen under no additional pressure), group 7 (low-temperature preservation at 0degrees C under pressure of 2 MPa), group 8 (low-temperature preservation at 0degrees C under no additional pressure), group 9 (low-temperature preservation at -5degrees C under pressure of 90 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in 37degrees C water bath, then MTT assay, WST-1 assay were processed. One way ANOVA and Tukey HSD method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization. In both MTT and WST-1 assay, group 7 (0degrees C/2 MPa) showed higher viability of periodontal ligament cells than other group (2-6, 8) and this was statistically significant (p < 0.05), but showed lower viability than group 1, immediate extraction group (no statistical significance). By the results of this study, low-temperature preservation at 0degrees C under pressure of 2 MPa suggest the possibility for long term preservation of teeth.
Subject(s)
Animals , Female , Humans , Rats , Anesthesia , Baths , Cryopreservation , Dimethyl Sulfoxide , Eosine Yellowish-(YS) , Freezing , Molar , Nitrogen , Periodontal Ligament , Tiletamine , ToothABSTRACT
This study examined the influence of the storage methods on the viability of oral epithelial cells using conventional cell freezing storage, slow freezing preservation, rapid freezing preservation, and slow freezing preservation with a pressure of 2 Mpa or 3 Mpa. The cell viability was evaluated by cell counting, WST-1 and the clonogenic capacity after 6 days of freezing storage. After 6 days, the frozen cells were thawed rapidly, and the cell counting, WST-1, and clonogenic capacity values were measured and compared. 1. The results from cell counting demonstrated that conventional cryopreservation, slow freezing under a 2 Mpa pressure and slow freezing under a 3 Mpa pressure showed significantly higher values than slow freezing preservation and rapid freezing preservation (p<0.05). 2. The results from the optical density by WST-1 demonstrated that slow freezing under a 2 Mpa pressure showed significantly higher values than slow freezing preservation and rapid freezing preservation (p<0.05). 3. The clonogenic capacity demonstrated that slow freezing under a 2 Mpa pressure showed significantly higher values than slow freezing preservation and rapid freezing preservation (p<0.05).
Subject(s)
Cell Count , Cell Survival , Cryopreservation , Epithelial Cells , FreezingABSTRACT
We report in detail ultrastructural changes and freezing damage mechanism about heart muscle and some organelles after rapid freezing. The ventricles of rat heart were cut pieces about 100-150/?m by microsiicer. The pieces were quickly injected liquid cryogen Freon 22 by Reichert-Jung spring-assistant mechanism (KF-80). The specimens frozen were rapidly transferred into substitution medium aceton and kept at -80℃(28h), then -60℃ (48h),-20℃(12h)and 4℃ (1 h). The structures of specimens frozen were well and there were no ice crystals in the area of the tissue frozen surface to 20?m depth. However, there were freezing damages in mitochondrial crista, intercellular substance and muscular fibre in the tissue surface to 30?m depth. The structure of tissue was destroyed by ice crystal over 50?m depth in the tissue. The results suggest that intercellular substance and mitochondrial crista are the most sensitive to ice crystal damage after rapid freezing of heart tissue, then the less sensitive are muscular fibre and nucleus. The unit membrane is not easy to be damaged by ice crystal.
ABSTRACT
This paper reports the effects of the temperature control freezing and the liquid nitrogen vapour freezing on the viability of rabbit morulae. After rapid thawing, 70.27% embryos were able to develope well in vitro in the temperature control freezing group and in the liquid nitrogen vapour freezing group, the developmental rate was 80.56% when the freezing medium contained higher concentration of glycerol. After synchronous embryos were transplanted, the birth rate was 13.64% and 13.33%, respectively, in both freezing groups, and 11.11% in the control group. The results indicate that both freezing methods have almost the same good results, but the method of liquid nitrogen vapour freezing is more useful than the temperature control freezing method in some aspects.