Objective:To investigate the molecular mechanism of rabbit ear cartilage remodeling induced by 1064 nm Nd∶YAG laser irradiation.Methods:Thirty-three rabbits were divided into 6 groups according to a random number table, and were used in the following experiments respectively: screening the optimal laser energy (6 rabbits), observing the changes of ear cartilage tissue immediately (6 rabbits), 1 week (6 rabbits), 3 weeks (6 rabbits), and 6 weeks after irridiation (6 rabbits), as well as transcriptome sequencing and verification (3 rabbits). The left ears of rabbits were used as untreated self-controls, while the right ears were irradiated. (1) Laser energy screening: the right ear cartilages of 6 rabbits were irradiated with 1064 nm Nd∶YAG laser at different energy densities (50, 60, 70, 80, 90, 100 J/cm 2). Cartilages without skin and perichondrium were cut into the same thickness and size for HE staining, in order to observe the changes of chondrocytes and to determine the optimal laser energy density for shaping (hereinafter referred to as the optimal energy density). (2) Histological observation of rabbit ear cartilage: the right ears of 24 rabbits were irradiated with laser under the optimal energy density. Bilateral cartilage samples were collected immediately after surgery and at 1, 3 and 6 weeks after surgery for histological observation (HE, Masson and Sirius red staining). (3) Transcriptome sequencing and sample verification: the right ears of 3 rabbits were irradiated with laser under the optimal energy density, and the cartilages were harvested and mixed within 6 hours after irradiation, which was set as the laser irradiation group. Cartilages of the same region and size on the left ears were set as blank control group. Transcriptome RNA sequencing was performed, and the expression levels of related genes and proteins were detected by quantitative polymerase chain reaction (qPCR) and Western blotting. Results:(1) After laser irradiation with different energy density of 50-100 J/cm 2, HE staining showed that the chondrocytes could be changed at the laser energy density of 80 J/cm 2, without causing vacuolar deformation and coagulation necrosis, indicating that the damage degree was suitable.(2) After 80 J/cm 2 laser irradiation, histological observation revealed that there was an irradiated zone immediately after the operation. The overall morphology of chondrocytes in the radiation zone was elongated and exhibited spindle cell-like changes, with deep matrix staining and obvious refraction, as well as a relative increase of type Ⅱ collagen. At 1 week after irradiation, the radiation zone became shallower and the cell size recovered. From 3 weeks to 6 weeks, the matrix staining around the radiation zone gradually deepened and the overall cellular morphology was stretched again. (3) Transcriptome RNA sequencing revealed that there were 198 differentially expressed genes (70 up-regulated and 128 down-regulated) in the laser irradiated group compared with the blank control group. Through further screening and study, CREB3L2 gene expression was up-regulated in the laser irradiation group. qPCR results showed that the relative expression level of CREB3L2 mRNA in laser irradiation group was significantly higher than that in the blank control group, the difference was statistically significant( P<0.01). Western blotting showed that the relative expression level of CREB3L2 protein was higher than that of control group in a time-dependent manner, and the difference was statistically significant( P<0.01). Conclusions:After irradiation with long-pulsed 1064 nm Nd∶YAG laser, the expression level of CREB3L2 gene is up-regulated, which then induced chondrocyte rearrangement and proliferation, resulting in morphological and biomechanical changes of ear cartilage.

Objective:To investigate the molecular mechanism of rabbit ear cartilage remodeling induced by 1064 nm Nd∶YAG laser irradiation.Methods:Thirty-three rabbits were divided into 6 groups according to a random number table, and were used in the following experiments respectively: screening the optimal laser energy (6 rabbits), observing the changes of ear cartilage tissue immediately (6 rabbits), 1 week (6 rabbits), 3 weeks (6 rabbits), and 6 weeks after irridiation (6 rabbits), as well as transcriptome sequencing and verification (3 rabbits). The left ears of rabbits were used as untreated self-controls, while the right ears were irradiated. (1) Laser energy screening: the right ear cartilages of 6 rabbits were irradiated with 1064 nm Nd∶YAG laser at different energy densities (50, 60, 70, 80, 90, 100 J/cm 2). Cartilages without skin and perichondrium were cut into the same thickness and size for HE staining, in order to observe the changes of chondrocytes and to determine the optimal laser energy density for shaping (hereinafter referred to as the optimal energy density). (2) Histological observation of rabbit ear cartilage: the right ears of 24 rabbits were irradiated with laser under the optimal energy density. Bilateral cartilage samples were collected immediately after surgery and at 1, 3 and 6 weeks after surgery for histological observation (HE, Masson and Sirius red staining). (3) Transcriptome sequencing and sample verification: the right ears of 3 rabbits were irradiated with laser under the optimal energy density, and the cartilages were harvested and mixed within 6 hours after irradiation, which was set as the laser irradiation group. Cartilages of the same region and size on the left ears were set as blank control group. Transcriptome RNA sequencing was performed, and the expression levels of related genes and proteins were detected by quantitative polymerase chain reaction (qPCR) and Western blotting. Results:(1) After laser irradiation with different energy density of 50-100 J/cm 2, HE staining showed that the chondrocytes could be changed at the laser energy density of 80 J/cm 2, without causing vacuolar deformation and coagulation necrosis, indicating that the damage degree was suitable.(2) After 80 J/cm 2 laser irradiation, histological observation revealed that there was an irradiated zone immediately after the operation. The overall morphology of chondrocytes in the radiation zone was elongated and exhibited spindle cell-like changes, with deep matrix staining and obvious refraction, as well as a relative increase of type Ⅱ collagen. At 1 week after irradiation, the radiation zone became shallower and the cell size recovered. From 3 weeks to 6 weeks, the matrix staining around the radiation zone gradually deepened and the overall cellular morphology was stretched again. (3) Transcriptome RNA sequencing revealed that there were 198 differentially expressed genes (70 up-regulated and 128 down-regulated) in the laser irradiated group compared with the blank control group. Through further screening and study, CREB3L2 gene expression was up-regulated in the laser irradiation group. qPCR results showed that the relative expression level of CREB3L2 mRNA in laser irradiation group was significantly higher than that in the blank control group, the difference was statistically significant( P<0.01). Western blotting showed that the relative expression level of CREB3L2 protein was higher than that of control group in a time-dependent manner, and the difference was statistically significant( P<0.01). Conclusions:After irradiation with long-pulsed 1064 nm Nd∶YAG laser, the expression level of CREB3L2 gene is up-regulated, which then induced chondrocyte rearrangement and proliferation, resulting in morphological and biomechanical changes of ear cartilage.

Objective:To summarize the experience of Bacillus Calmette-Guerin(BCG) in the treatment of bladder cancer secondary to renal transplantation.Methods:The clinical data of 5 patients who underwent BCG bladder irrigation after secondary bladder cancer after kidney transplantation in Tianjin First Central Hospital from January 2015 to December 2019 were analyzed. There were 1 male and 4 female cases. During the period of immunosuppression after transplantation, 1 case developed secondary high-level non-muscular invasive bladder cancer (NMIBC), 3 cases developed secondary low-grade NMIBC, and 1 case developed secondary glandular cystitis (4 cases). The mean age of the 5 patients with secondary bladder cancer was 59.7±4.0 years. Case one with high level NMIBC was treated with transurethral resection of bladder tumor (TURBT) and postoperative irrigation of epirubicin. Case 3 and 5 with low-level NMIBC accepted regular postoperative irrigation of gemcitabine. No irrigative therapy was performed in case 2. Bladder cancer recurred in case 1, 2, 3 and 5 after 20.1±9.7 months. TURBT was observed in all the 4 patients, among which 3 were of high grade NMIBC and 1 was of low grade NMIBC. Four patients were irrigated with BCG 2 weeks after operation. Postoperative pathology indicated low-level NMIBC in case 4, and BCG was irrigated 2 weeks after the operation. During perfusion therapy, immunosuppressive agents were continued.Results:During BCG perfusion, 4 of the 5 cases showed BCG related local inflammation, among which 2 cases presented symptoms of bladder irritation, 1 case presented hematuria, and 1 case presented hematuria with low fever. Patients with frequent urination, pain in urine, hematuria and other symptoms improved after drinking plenty of water, taking bed rest and taking levofloxacin (0.5g/ day ×7 days). Patients with low fever were treated with antipyretic treatment. No antituberculous agents were used prophylactically during BCG perfusion. There were no symptoms of tuberculosis infection or sepsis. The function of transplantated kidney was normal and no tendency of rejection. The 5 patients were followed up for 7-24 months, 1 patient was lost to follow-up after 7 months of BCG bladder perfusion, and no tumor recurrence or metastasis was found in 5 patients during the follow-up.Conclusions:The use of immunosuppressive agents does not reduce the biological activity of BCG, and BCG does not increase the risk of systemic toxicity or affect the function of transplanted kidneys in immunocompromised patients. BCG is a treatment option for bladder cancer secondary to renal transplantation.

Objective:To investigate the 2 years’ efficacy of intravesical instillation of domestic BCG versus epirubicin in the prevention of recurrence of intermediate-risk or high-risk non-muscular invasive bladder cancer and predictive factors of BCG instillation.Methods:From July 2015 to June 2020, 18-75 years old patients with moderate to high-risk non muscle invasive bladder cancer (NMIBC) confirmed by pathological examination were involved. The ECOG score was 0-2. Exclusion criteria included ①immune deficiency or impairment (such as AIDS), using immunosuppressive drugs or radiotherapy, suspected allergic to BCG or epirubicin or excipients of the two drugs, fever or acute infectious diseases including active tuberculosis or receiving anti tuberculosis treatment, with severe chronic cardiovascular and cerebrovascular diseases or chronic kidney disease; ②combined with other urogenital system tumors or other organ tumors; ③combined with muscle invasive bladder urothelial carcinoma (≥T 2); ④undergoing chemotherapy, radiotherapy or immunotherapy within 4 weeks (immediate instillation after surgery not included); ⑤ pregnant or lactating women; ⑥ comfirmed or suspected bladder perforation; ⑦gross hematuria; ⑧cystitis with severe bladder irritation that may affect the evaluation; ⑨participat in other clinical trials within 3 months; ⑩alcohol or drug addiction; ?any risk factors that may increasing the risk of patients. Epirubicin 50 mg was irrigated immediately after the operation(TURBT or laser resection). The patients were randomly divided into BCG15 group, BCG19 group and epirubicin group by the ratio of 2∶2∶1, and the patients were maintained intravescical instillation for 1 year. The recurrence and adverse events of the three groups were compared. Univariate and multivariate analysis was performed to predict the risk factors of BCG irrigated therapy failure. Result:By June 15, 2020, the median follow-up duration was 22.1 months(12.1, 32.3), and there was no statistical difference between the groups ( P=0.9024). There were 274 patients enrolled in BCG19 group, 277 patients enrolled in BCG15 group and 130 patients enrolled in the epirubicin group. The drop-off rate was 16.6%(113 cases)and made no difference between groups( P=0.6222). There were no significant difference in age, gender, BMI, or ECOG score( P>0.05). During the follow-up, 116 cases was detected recurrence or progression. The recurrence rate of the three groups was 14.2% and 14.8% in BCG19 group and BCG15 group, and 27.7% in the epirubicin group. There was no difference in recurrence rate between BCG19 and BCG15 group( P=0.9464). The recurrence rate of BCG19 group was lower than that of the epirubicin group ( P=0.0017). The recurrence rate of BCG15 group was lower than that of the epirubicin group ( P=0.0020). There was no difference in the cumulative recurrence free survival rate between BCG19 and BCG15 group (95% CI0.57-1.46, P=0.7173). The cumulative recurrence free survival rate of BCG 19 group was better than that of the epirubicin group( HR=0.439, 95% CI0.26-0.74, P=0.0006), and the cumulative recurrence free survival rate of BCG15 group was better than that of the epirubicin group ( HR=0.448, 95% CI0.29-0.80, P=0.0021). The total incidence of adverse events in 19 BCG19, BCG15 and epirubicin group were 74.5%, 72.6% and 69.8% respectively. There was no difference in the incidence of adverse events between BCG19 and BCG15 group( P=0.6153). The incidence of adverse events in epirubicin group was lower than that of BCG19( P=0.0051) and BCG15( P=0.0167) groups.There was no significant difference in the incidence of serious adverse events (SAE) among the three groups ( P=0.5064). Log rank test univariate analysis and Cox risk regression model multivariate analysis showed that the history of bladder cancer recurrence( HR=6.397, 95% CI1.95-20.94, P=0.0001)was independent risk factor for BCG irrigation failure. Conclusions:The 2 years’ efficacy of intravesical instillation of domestic BCG is better than than of epirubicin with good tolerance and safety. There is no difference between BCG19 and BCG15 group. BCG doesn’t increase SAE compared with epirubicin. Recurrence status was an independent prognostic factor regarding recurrence-free survival.

Objective To explore the potential role of BK virus (BKV) in urothelial carcinoma after renal transplantation .Methods From May 2013 to March 2017 , We collected 26 cases of urothelial carcinoma after renal transplantation (study group) and 30 cases of urothelial carcinoma of non-immunosuppressed patients (control group) .Tumor tissues were stained with SV40 T antigen by immunohistochemical assay .Urinary and peripheral blood samples were assayed for BKV-DNA levels by real-time fluorescent quantitative polymerase chain reaction (PCR) .Results There were 7 positive cases of SV40 T-Ag in study group and only 1 weakly positive case in control group .The positive rate of BKV-DNA was 38 .5％ in urinary samples in study group (10 /26 ) and it was significantly higher than that in control group by 10％ (3/30)(P< 0 .05) .And the positive rate of blood BKV-DNA was not different between two groups ( P > 0 .05 ) .Conclusions There is a relatively high prevalence of BKV infection in urothelial carcinoma after renal transplantation .And a ,high level of BKV infection may play a role in urothelial carcinoma after renal transplantation .

Objective To investigate the expression of human leucocyte antigen G (HLA-G) in urothelial carcinoma after renal transplantation,and to analyse the relationship between HLA-G expression and the various clinical and pathological parameters.Methods 29 patients with urothelium carcinoma after renal transplantation for the first time from January 2005 to June 2016 were selected as the experimental group,the age range was 32-70 years,with an average of (55.5 ± 8.1) years.29 non-transplanted patients with urothelial carcinoma as the control group 1,the age range was 36-74 years,with an average of (57.9 ± 8.2) years.15 cases of normal urinary tract epithelial were from cystoscopy biopsy as the control group 2.Immunohistochemical method was used to detect the difference of HLA-G expression between the three groups.The clinical and pathological data of patients with urothelial carcinoma after renal transplantation were analyzed.Results The expression rate of HLA-G was 79.3％ (23/32) in patients with urothelial carcinoma after renal transplantation,37.9％ (11/32) in non-transplanted group and 0 (0/15) in normal urinary tract epithelium group.The expression rate of HLA-G in non-transplanted group was significantly higher than that in normal urinary tract epithelium group (P ＜ 0.05).The expression rate of HLA-G in patients with urothelial carcinoma after renal transplantation was significantly higher than that in nontransplanted group and normal urinary tract epithelium group (P ＜ 0.05).Conclusions HLA-G is associated with the occurrence of urothelial carcinoma after renal transplantation.It may provide a new idea for the prevention and treatment of urinary tract epithelium after renal transplantation.

OBJECTIVETo observe the effect of different doses of acetamide on the histopathology in the cerebral cortex of rats with tetramine (TET) poisoning and to provide a basis for the treatment of fluoroacetamide poisoning with acetamide.

METHODSEighty clean Sprague-Dawley rats were randomly divided into five groups: saline control group,dimethylsulfoxide water solution control group,TET poisoning group, acetamide (2.88 g/kg/d) treatment group, and acetamide (5.68 g/kg/d) treatment group, with 16 rats in each group. Rats in the poisoning group and treatment groups were poisoned with TET by intragastric administration after fasting; then, saline was injected intramuscularly into rats of the poisoning group, and different doses of acetamide were injected intramuscularly into rats of treatment groups; the course of treatment was 5 d. At 3 h, 12 h, 48 h, and 7 d after treatment, the cerebral cortex was harvested from rats in each group, and the histopathological changes in the cerebral cortex were evaluated under light and electron microscopes.

RESULTSThe light microscopy showed that the TET poisoning group had hypoxia changes in the cerebral cortex, which worsened over time; the treatment groups had reduced hypoxia changes, and the acetamide (2.88 g/kg/d) treatment group had more reduction than the acetamide (5.68 g/kg/d) treatment group. The electron microscopy showed that the apoptosis of neuronal cells were the main pathological changes in the TET poisoning group; the treatment groups had reduced apoptotic changes, and the acetamide (2.88 g/kg/d) treatment group had more reduction than the acetamide (5.68 g/kg/d) treatment group.

CONCLUSIONNo pathological changes associated with the synergistic toxic effect of acetamide and TET are found in the cerebral cortex. Acetamide (2.88 g/kg/d) could reduce central nervous lesions, but the efficacy is not improved after increasing the dose. For patients who cannot be identified with TET or fluoroacetamide poisoning, acetamide could be considered for treatment.

Acetamides ; pharmacology ; Animals ; Bridged-Ring Compounds ; toxicity ; Cerebral Cortex ; drug effects ; pathology ; Disease Models, Animal ; Male ; Rats ; Rats, Sprague-Dawley

Objective To compare healing effects of surgical incisions using 635 /808 nm dualwavelength semiconductor laser and He-Ne laser irradiation.Methods 168 cases of non-malignant tumor surgery patients were randomly divided into 2 groups:test group including 83 cases which were treated on the surgical incisions by laser irradiation of the semiconductor illumination with low-intensity power; positive control group with 85 cases treated with He-Ne laser.Observation was carried out on incision healing by the clinic manifestations including redness,heat,swelling,pain,exudation,wound open,adverse event and the incision length offset.Results There was no significant difference between experimental group and the control group on incision healing (P＞0.05).Conclusions The treatment efficacy of the dual-wavelength semiconductor laser on surgical incisions healing is similar with that of the He-Ne laser.

Objective To evaluate the safety,feasibility and results of the hand-assisted retroperitoneal laparoscopic living donor nephrectomy ( HRPLDN ) with a modified technique. Methods Living donors (n =32) were divided into HRPLDN group (n =16) and open group (n =16) according to surgical technique.Operative data and postoperative outcomes including operative time,estimated blood loss,warm ischemia time,length of hospital stay and complication rate,were collected. Results All procedures were completed successfully.In HRPLDN group,the mean operative time was 101.3 ± 21.2 min (range from 70 to 150 min),with an estimated blood loss of 53.8 ±25.5 ml (range from 20 to 100 ml) and warm ischemia time of 2.4 ± 0.6 min ( range from 1.5 to 3.5 min).No living donor needed conversion to open surgery and the urine volume of transplanted kidney after first 24 hours was 5036 ml (range from 3500 -6500 ml).The mean postoperative on bed time were (2.8 ± 0.7 ) d (ranging from 2 -4 d).All parameters of HRPLDN were significantly better than that of open groups. Conclusion Living donor nephrectomy with HRPLDN is a safe and reliable surgical technique.

Objective To analyze the incidence and clinical features of urothelial tumors in renal allograft recipients.Methods A retrospective analysis of 1042 patients received renal allografts who had taken immunosuppression for at least six months between 2006 and 2011 in The First Centre Hospital of Tianjin was performed.Results Eleven cases of uroepithelial tumors were diagnosed in the 1042 cases of renal transplantation ( 1.06％ ),of whom 9 cases were noticed by hematuria ( 81.8 ％ ),2 cases ( 18.2％ ) by medical examination.Six patients were diagnosed with multifocal urothelial carcinomas.Surgery was performed on all the patients with renal tumors and followed by chemotherapy or radiotherapy.Conclusion Malignancies in urinary tract after renal transplantation should be bore in mind.Early diagnosis is very important.The treatment options include reducing immunosuppressive agents and removing tumor lesions completely.