Objective To investigate the effects of ?-aescin on the germ cell apoptosis and expression of bcl-2 and Bax proteins in bilateral testes following ipsilateral testicular torsion and detorsion. Methods Thirty adolescent male Wistar rats were randomly divided into control group (C group), model group of testicular torsion and detorsion (M group) and ?-aescin treatment group (T group), each group containing 10 animals. In M and T groups, rats underwent a clockwise 720? ipsilateral testicular torsion for 2 h followed by detorsion, and the rats in T group were intraperitoneally injected with ?-aescin 15 min before detorsion. 48h after operation, bilateral testes were collected, and used to detect cell apoptosis and expressions of bcl-2 and Bax by TUNEL technique and immunohistochemical method, respectively. Results In M group, the apoptotic index of the germ cells and Bax expression of bilateral testes significantly increased compared with C group (P

Objective To compare efficacy, time consumed and complications among cricothyroid membrane puncture guided tracheostomy ( CMPGT ) , surgical tracheostomy ( ST ) , surgical cricothyroidotomy ( SC) , and percutaneous tracheostomy using Griggs'guide wire dilating forceps ( GWDF) for the establishment of airway in urgent need of medical attention.Methods Twenty miniature swine were randomly ( random number) divided into four groups.The procedures of CMPGT, ST, SC and GWDF were carried out when patients'SpO2 ( oxygen saturation of blood ) declined to 80% by suspension of oxygen supply after general anesthesia. Procedure performed time, ventilation resumed time, SpO2 and electrocardiograph ( ECG) and arterial blood gases ( ABG) analysis including SaO2 , PaO2 , PaCO2 , blood pH, and heart rate, blood pressure were recorded.Fiberoptic bronchoscope was used to assess any damage to the tracheal wall.Complications were noted and scored in a two-month follow-up period.Results Airways were successfully established in all swine.The times consumed for SC, GWDF, CMPGT, and ST were (86 ±12) s, (165 ±63) s, (174 ±34) s, and (519 ±128) s, respectively, however a shorter time for ventilation resumed was found in CMPGT procedure (23 ±4) s, P<0.01.ECG showed that SpO2 and T-wave decreased and Q-T shortened after oxygen suspension and recovered to normal level rapidly after ventilation.There were significant differences in ECG and ABG between pre-and post-operative periods ( P<0.05) in all groups.Minimal intra-operative bleeding was found in two swine of each group.In ST group, moderate intra-operative bleeding was encountered in three swine.Three pigs were suffered from hypotension owing to prolonged hypoxemia.There was minimal postoperative bleeding occurred in one swine, thus leading to stoma infection.In SC group, moderate intra-operative bleeding was noticed in one swine. One miniature swine had slight injury at laryngeal cartilage resulting in difficult decannulation happened in one swine.and moderate tracheal wall injury occurred in one swine of GWDF group.The complication scores of CMPGT, GWDF, SC and ST were 3, 5, 9, and 19, respectively.Conclusions A shorter time for ventilation resumed with fewer complications was found in the procedure of CMPGT than that in the other methods.This animal study suggested that CMPGT was a time-saving and secure technique for emergency surgical airway establishment which has been practiced in human being as well.Key words:Surgical airway establishment, Cricothyroidotomy, Tracheostomy, Percutaneous dilational tracheostomy.

BACKGROUND:In 2001, Zuk et al found adipose-derived stem cels (ASCs) from the aspirate of liposuction for the first time, which launched a new era of stem cel research. In recent years, stem cels have been proved to widely exist in many tissues and organs. ASCs are always in the spotlight of plastic and reconstructive surgery, tissue engineering and regenerative medicine because of extensive sources and simple isolation.
OBJECTIVE: To review the fat tissue harvesting and ASCs isolation, purification, expansion, and cryopreservation, to discuss the main factors which influence the yield, proliferation capacity and differentiation potential of ASCs, and to predict the future research interests based on current issues.
METHODS:On September 10th, 2015, relevant articles were searched in PubMed using the folowing format: (adipose stem cels[Title]) OR (adipose-derived stem cels[Title]) OR (adipose-derived mesenchymal stem cels[Title]) and in SinoMed using the folowing format in Chinese: (“adipose-derived stem cels” [Title])or(“adipose-derived mesenchymal stem cels”[Title]). Finaly, 81 representative articles were included according to their titles and abstracts. In this review, we also introduced relevant experience about the aforementioned procedures from the Department of Plastic Surgery and Tissue Regeneration and Molecular Cel Engineering Lab of University of Texas, MD Anderson Cancer Center, USA.
RESULTS AND CONCLUSION:The widely dispersed fat tissues potentialy provide abundant stem cels for tissue engineering and regenerative medicine. Liposuction is a mini-invasive approach for harvesting fat tissues. Colagenase digestion is the major method for ASCs isolation due to its simplicity and high yield in basic research. However, clinical fat transplantation without ASCs isolation or non-colagenase isolation of stromal vascular fraction or ASCs is preferred. The phenotype, proliferation and differentiation capacity of ASCs may be affected by several factors during the fat tissue harvesting and ASCs isolation. Therefore, a standard protocol for ASCs isolation is needed.

Objective Gambogic acid ( GA) can suppress the growth of multiple tumor cells , including gastric carcinoma , hepatoma , hematologic neoplasms and breast carcinoma , but there have been few reports about its effect on urologic neoplasms .This study was to investigate the possible mechanisms of GA inducing bladder cancer cell apoptosis and cell cycle arrest . Methods We cultured human bladder cancer BIU8-7 cell lines in vitor and treated the cells in the logarithmic growth phase with isotonic saline solu-tion (negative control)or GA at the concentrations of 1.0, 2.0, and 3.0μmol/L, respectively.We determined the expression of the Caspase-3 protein in the tumor tissue using the immunohistochemical S-P method and detected GA-induced apoptosis of the bladder cancer cells and cell cycle changes by flow cytometry . Results The expressions of the Caspase-3 protein were 4.28 ±1.86, 5.03 ± 0.78, and 6.47 ±1.31 in the 1.0, 2.0, and 3.0μmol/L GA groups, respectively, significantly higher than 2.13 ±1.27 in the nega-tive control (P<0.05).Flow cytometry showed a gradual decrease of the cells in the G 0/G1 phase and a gradual increase in the G2/M phase , but no obvious change in the S phase . Conclusion Gambogic acid can promote the apoptosis , arrest the cell cycle , and in-hibit the proliferation of bladder cancer cells by increasing the expression of the Caspase -3 protein.

Objective:To explore the changes of immune microenvironment and prognosis of bladder cancer patients with positive urinary nuclear matrix protein 22 (NMP22).Methods:Retrospective analysis was made on 86 patients who were diagnosed with bladder cancer in Xuzhou Central Hospital from January 2019 to September 2020. All patients were tested for urinary NMP22 by colloidal gold method. The patients with positive test results were NMP22 positive group, and the patients with negative test results were NMP22 negative group. The expression of CD8, programmed cell death-ligand 1 (PD-L1), programmed cell death protein-1 (PD-1) and PanCK were detected by multiple fluorescent immunohistochemical method on the pathological tissue sections of all enrolled patients with bladder cancer after surgery. Follow-up data of enrolled patients were collected after discharge, and univariate and multivariate Cox analysis was performed on the follow-up data.Results:There were 69 patients in the NMP22 positive group and 17 patients in the NMP22 negative group. The percentage of CD8 and PD-L1 positive cells in NMP22 positive group was significantly higher than that in NMP22 negative group, and the difference was statistically significant (all P<0.05). Univariate analysis showed that tumor stage was correlated with bladder cancer progression ( HR=2.67, P=0.017). Multivariate analysis showed that positive NMP22 was significantly correlated with bladder cancer recurrence and disease progression (all P<0.05). Conclusions:The density of CD8 + T cells and PD-L1 in tumor parenchyma of urinary NMP22 positive bladder cancer patients was higher than that of NMP22 negative patients. Urinary NMP22 positive can be one of the bad prognostic factors of bladder cancer, and the patients with NMP22 positive should strengthen reexamination.

Professor's experience is introduced in insomnia treated with the acupuncture technique for regulating the governor vessel and calming down the mind.'s acupuncture technique is in one of the series of character techniques in the acupuncture academic school in the region of Hebei province. Professorbelieves that insomnia resultes from the disharmony of the nutrient and the defensive and the dysfunction of spleen and stomach. The acupoints are selected on the basis of regulating the governor vessel, calming down the mind, nou-rishing the water and the wood to harmonize the mind, strengthening the spleen and stomach to calm down the mind, in which the acupoints of the governor vessel and theprimary points are used in combination. For regulating the governor vessel, Baihui (GV 20) and Shenting (GV 24) are used, combined with Zhongwan (CV 12) for regulating the nutrient and the defensive and calming down the mind. The-primary points of the heart, liver and kidney meridians are selected to nourish water and wood and harmonize the mind. Tianshu (ST 25), Zhongwan (CV 12) and Yinlingquan (SP 9) are used to strengthen the spleen and stomach and for the treatment of the root cause of disease. Additionally, the sequence of needling, the needling techniques for reinforcing and reducing, as well as needle withdrawing are considered. The idea of regulating the governor vessel and calming down the mind as well as the needling techniques for insomnia are considerably introduced in the paper.

Objective To study the effect and mechanism of Momordica anti-HIV protein of 30 ku (MAP30) on the migration of bladder cancer.Methods The IC50 of human bladder cancer 5637 and T24 cells was calculated by methyl thiazolyl tetrazolium (MTT) method.The migration ability of these two cells was evaluated by scratch migration test and Transwell cell migration test.The expression of migrating proteins such as matrix metalloproteinases (MMPs) and adhesion molecule N-cadherin were compared by Western blot.Results Scratch migration test:there were significant differences in migration rates of 5637 cells at 8 h and 22 h (P ＜ 0.05).There were significant differences in migration rates of T24 cells at 22 h (P ＜ 0.05),but no significant differences in migration rates at 8 h (P ＞ 0.05).The expression of Vimentin,Fibronectin,MMP-2,MMP-9 and N-Cadherin in 5637 cells and T24 cells of human bladder cancer decreased significantly after adding MAP30.The E-Cadherin expression in human bladder cancer 5637 cells were decreased,but no target band was detected in human bladder cancer T24 cells.Conclusions The ribosome-inactivating protein MAP30 can effectively inhibit the migration of human bladder cancer 5637 and T24 cells by inhibiting the EMT pathway and inhibiting the expression of MMPs.

Objective To study the effect of enhancer of rudimentary homolog (ERH) gene on migration and invasion in human bladder cancer T24 and 5637 cells.Methods After knocking out the ERH gene of human bladder cancer T24 and 5637 cells,Wound healing assay,Transwell cell migration assay and Transwell cell invasion assay were used to verify the migration and invasion function.Cell migration related protein was detected by Western blot.Nude mouse tail vein transfer assay was used to study the metastasis ability of bladder cancer cells in vivo.Results (1) The Wound healing assay showed that there were significant differences in the migration cell counts of human bladder cancer 5637 and T24 (P ＜ 0.05).(2) There were significant differences in migration and invasion cell counts of Transwell assay (P ＜0.05).(3) Western blot showed that the expression of E-Cadherin in human bladder cancer 5637 cells and T24 cells was significantly increased (P ＜ 0.05) after knocking out ERH gene,while the expression of Fibronectin,Twist,Vimentin and Snail2 protein were significantly decreased (P ＜ 0.05).(4) Nude mouse tail vein transfer assay showed that lung metastases were significantly reduced in the ERH knockout group compared with the normal ERH group.Conclusions Both in vitro and in vivo experiments suggest that ERH knockout affects the migration and invasion of human bladder cancer T24 and 5637 cells.

Objective To investigate the effect of ERH gene knockdown on the proliferation and apoptosis of human bladder cancer T24 cells. Methods T24 cells infected by lentivirus with interference on ERH gene sequence were cloned to establish stable T24 cells clone in ERH gene suppression. The expression of ERH mRNA gene in bladder cancer was detected by using quantitative real time polymerase chain reaction (qPCR). The effects of ERH knockout on the cell proliferation and apoptosis were examined by using methylthiazolyl tetrazolium (MTT) assay, colony formation assay and flow cytometry. The effect of ERH knockout on the tumorigenic effect of T24 cells in vivo was verified by subcutaneous tumor formation in nude mice. Results After lentiviral transfection, qPCR results showed that the knockdown effect of ERH mRNA in ERH normal group (untreated T24 cells) was better than that in ERH gene knockdown group, and the difference was statistically significant [(1.006±0.126) vs. (0.079±0.007); t=12.72, P=0.0002]. After knocking out ERH gene, MTT assay showed that the proliferation ability of T24 cells in ERH gene knockdown group was weakened compared with ERH normal group, and the difference was statistically significant [A490 value: (0.13±0.00) vs. (0.66±0.01);t=104.61, P<0.0001]. Colony formation assay indicated that the ability of clone in ERH normal group was weakened compared with ERH gene knockdown group [(10.5 ±1.2) vs. (196.4 ±4.0); t= 73.63, P< 0.0001]. Flow cytometry showed that the cell apoptosis rate in ERH gene knockdown group was higher than that in ERH normal group [(11.0 ±0.5) ％ vs. (4.2 ±0.5) ％; t= 16.06, P<0.0001]. Imaging results of subcutaneous tumor formation in nude mice showed that the total fluorescence intensity of the tumor area in ERH gene knockdown group was (4.67 ±0.59) × 1010 μW/cm2, and the corresponding part in ERH normal group was (9.54±4.20) × 1010μW/cm2 (t=3.64, P=0.0051);tumor weight in ERH gene knockdown group was (0.80±0.62) g, and in ERH normal group was (1.79±0.71) g (t=3.33, P=0.0037). Conclusion ERH gene knockout can inhibit the proliferation of human bladder cancer T24 cells, and promote the cell apoptosis.

Objective:To investigate the effect of 125I-RSOAds-hTERT/PSA oncolytic adenovirus on targeted therapy of prostate cancer and its effect on tumor microenvironment. Methods:125I-RSOAds-hTERT/PSA ( 125I-virus complex) oncolytic adenovirus was constructed by PCR amplification and double restriction enzyme ligation. TUNEL staining, flow cytometry and Caspase-3 immunoblotting assay were used to detect the killing effect of 125I-RSOAds-hTERT/PSA oncolytic adenovirus on prostate cancer cells in vitro and in vivo, respectively. To explore the effect of 125I-virus complex on tumor tissue cytokine secretion levels, interleukin 2 (IL-2), IL-10, tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in the culture supernatant of human prostate cancer cell line PC3, mouse prostate adenocarcinoma cell line RM-1, and mice serum were detected by ELISA. We explored the regulation of 125I-virus complex on the expression of CD24, CD44 and prostate stem cell antigen (PSCA) in prostate tumor tissues and tumor cells through immunohistochemistry. Meanwhile, the expression levels of CD32 and vascular endothelial growth factor (VEGF), as well as CD4+ , CD8+ and macrophage infiltration in tumor tissue were detected through immunofluorescence experiments. Results:125I-virus complex oncolytic adenovirus significantly increased tumor cell apoptosis in vitro and in vivo that was significantly higher than that of 125I group and virus complex group. Meanwhile, IL-2 ( t=-183.30, -38.20, P<0.05), IL-10 ( t=113.80, 92.71, P<0.05), TNF-α ( t=-73.20, -73.91, P<0.05), IFN-γ ( t=-65.37, -139.70, P<0.05) increased in vitro and in vivo. 125I-virus complex reduced the expression of CD24, CD44 and PSCA in tumor cells and tumor tissue, reduced the weight of tumor tissue, inhibited angiogenesis of tumor tissue ( t=8.55, P<0.05), and regulated the immune response in tumor tissue. Conclusions:125I-virus complex targeting prostate cancer can significantly kill cancer cells, reduce the weight and angiogenesis of tumor, and improve tumor microenvironment.