Aim To evaluate the effect of expression inhibition of spermine oxidase(SMO)on the actitumor activity of polyamine analogue CPENSpm (N~1-cyclopropylmethyl-N~(11)-ethylnorspermine).Methods siRNA technique was used to inhibit expression of SMO in human lung cancer line A549.QT-RT-PCR and enzyme activity assay was performed to determine the expression level of SMO.The cell proliferation was detected by MTT assay.The apoptosis of A549 cells were evaluated by DNA degradation and Sub-G_1/flow cytometry assay.Results The A549 cell line with silenced SMO expression was successfully obtained.Basic SMO mRNA and enzyme activity levels in the SMO-siRNA plasmid transfected cells were 0.53% and 14% lower than that in the control cells respectively. Treating A549 control cells by 10 μmol·L~(-1) CPENSpm for 24 hours resulted in a 10-folds up-regulation of SMO in mRNA level and 20-fold increase in enzyme activity,but this drug-induced SMO expression was obviously prevented in SMO-siRNA plasmid transfected cells.MTT assay demonstrated that SMO expression inhibition decreased the sensitivity of A549 cells to CPENSpm exposure(0～20 μmol·L~(-1)).DNA degradation and sub-G_1 assay proved a deceased ability of CPENSpm to induce apoptosis in SMO-siRNA plasmid transfected cells.Conclusion Up-regulation of SMO by CPENSpm is possibly one of the molecular basics for its antitumor activity.

AIM: To prepare recombinant human spermine oxidase (SMO) and polyclonal antibody against human SMO by gene recombination techniques. METHODS: Human SMO cDNA was amplified from total RNA of A549 cells through reverse transcription PCR. The cDNA was then cloned into pET-15b to construct SMO prokaryotic expression vector. After transforming, the vector was induced to express recombinant SMO by IPTG in E. coli BL21 (DE_3). Recombinant SMO was purified by Ni-NTA resin under denaturing condition and then was dialyzed to renature. The enzyme activity of recombinant SMO was analyzed by chemical fluorescent method. SMO polyclonal antibody was prepared by using recombinant human SMO protein purified by polyacrylamide gel electrophoresis as antigen to inoculate rabbit intradermally. The titer and specificity of anti-sera were determined by ELISA, Western blot and Immune Cell Chemistry. RESULTS: Purified and dialyzed recombinant human SMO has the specificicity of oxidizing the spermine. The polyclonal antibody has high titer and specificity against human SMO. CONCLUSION: This research established a method for prokaryotic expression, purification and polyclonal antibody preparation of human SMO. The method lays a foundation for the future functional research of SMO.

Objective To investigate whether difluromethylornithine (DFMO) can be used in the treatment of human leukemia. Methods The cell proliferation was detected by MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium)] assay after treatment of human lymphocyte Jurkat cells by DFMO (0 to 10 mmol/L) for 24 to 72 h. Enzyme activity of spermine oxidase (SMO) and acetylpolyamine oxidase (PAO) was determined by chemiluminesence assay. DNA fragmentation assay was used to evaluate cell apoptosis. Fluorescent dye assay was performed to determine the changes in mitochondrial membrane potential. Western blotting was used to determine Bax content. Casepase-3 enzyme activity was measured by spectrophotometric method. Results DFMO treatment inhibited the proliferation of Jurkat cells significantly in a dosage- and time-dependent manner (P

Objective To investigate the effectiveness and cost of 50% and 80% wettable powder of niclosamide ethanolamine salt (NESWP) and 26% metaldehyde and niclosamide suspension concentrate (MNSC) in hilly schistosomiasis-endemic regions, so as to provide insights into the selection of chemical molluscicides in hilly regions. Methods In September 2020, a wasteland in Guanshanqiao Village, Yanrui Township, Yushan County of Jiangxi Province was selected as the experimental region, which was sectioned into five blocks and defined as four experimental groups (A₁, A₂, B, C) and a blank control group (D). 80% NESWP were given at doses of 1 g/m² and 1.5 g/m² in groups A₁ and A₂ using the spraying method, 50% NESWP was given at a dose of 2 g/m² in Group B using the spraying method, and 26% MNSC was at a dose of 4 g/m² in Group C using the spraying method, while no chemical treatment was given in Group D. Snail survey was performed using a systematic sampling method before chemical treatment and 1, 3, 7 d and 15 d post-treatment to examine the molluscicidal effect, and all molluscicidal costs were estimated to calculate the cost of chemical treatment per 1 m² and the cost of the reduction in the mean density of living snails per 1%. Results The highest mortality of snails was 78.95% and the lowest density of living snails was 0.2388 snails/0.1 m² in the experimental groups within 7 d of chemical treatment, and the highest mortality of snails was 94.74% and the lowest density of living snails was 0.058 0 snails/0.1 m² 7 d post-treatment. There were no significant differences in the snail mortality among the A₁, A₂, B and C groups 1 (χ² = 2.250, P > 0.05), 3 (χ² = 1.779, P > 0.05) or 15 d post-treatment (χ² = 2.286, P > 0.05), while a significant difference was detected in the snail mortality among the four groups 7 d post-treatment (χ² = 7.990, P = 0.046). In addition, there were no significant differences in the snail mortality between A₁ and A₂ groups 1 (χ² = 0.724, P > 0.05), 3 (χ² = 0.584, P > 0.05), 7 (χ² = 0.400, P > 0.05) or 15 d post-treatment (χ² = 0.251, P > 0.05). The costs of chemical treatment per 1 m² were 0.58, 0.60, 0.64 Yuan and 0.73 Yuan in groups A₁, A₂, B and C, and the costs of the mean density of living snail per 1% reduction were 19.29, 20.44, 21.68 Yuan and 23.53 Yuan in groups A₁, A₂, B and C, respectively. Conclusion 80% NESWP shows a high molluscicidal efficacy and low cost in hilly schistosomiasis-endemic regions.

Background and purpose: Primary signet ring cell carcinoma(SRCC) of the bladder is rarely diagnosed in the clinic. Few cases have been reported in the literature, so there was lack of understanding of the primary bladder SRCC in terms of diagnosis and treatment. Our study was to investigate the clinical features and treatment strategy for primary SRCC of the bladder and review the status of the disease along with the literature. Methods: 3 cases of primary bladder SRCC were studied, including clinical features, treatment, follow-up and their prognosis.The literature was reviewed. Results: All cases received ultrasound, computerized tomography, cystoscopy, biopsy and other related lab tests for diagnosis and differential diagnosis. Laparoscopic radical cystectomy and orthotopic ileal neobladders were performed in 2 cases, while the other case received laparoscopic radical cystectomy and ileal conduit diversion, Chemotherapy (cisplatin and 5-fluorouracil) was delivered in one case after surgery. One patient died at 6 months postoperatively because of multiple metastasis. The other 2 cases have been followed-up only for 8 and 12 months postoperatively, and no recurrence or metastasis have been observed. Conclusion: Primary SRCC of the bladder lacks distinctive clinical and imaging manifestations. The tumor grows very invasively. Radical cystcctomy is one of the optimal approaches for treatment of SRCC of bladder.

Background and purpose: The prognostic factors on survival for the patients with prostate carcinoma are still underdeterrnined. This study was to analyze the survival of three common treatment methods for prostate carcinoma and the prognostic factors on survival. Methods: 494 male patients who were diagnosed as prostate cancer were enrolled into the retrospective study. All of the data like age, stage, grade, PSA level, ALP, Hb and treatments were collected. Overall survival and disease specific survival rates for patients were analyzed by Kaplan-Meier method. Prognostic factors on disease specific survival were also analyzed by Log-rank test and Cox proportional hazards model. Results: Disease specific survival rates at 1, 3 and 5 year were 96.0%, 89.0% and 80.0% for all 494 patients, respectively. Disease specific survival rate at 3-year was 92.4% for brachytherapy, 100.0% for radical prostatectomy and 80.6% for hormonal therapy (P=0.008). Multivariate analysis by Cox model showed that stage, PSA level and age significantly impacted on disease specific survival. Conclusion: Brachytherapy and radical prostatectomy provides longer survival time than hormonal therapy for patients with prostate cancer. Clinical stage and PSA level and age of prostate cancer are independent factors impacting on survival significantly.

Objective: To evaluate the long-term functional recovery after contralateral 7th cervical nerve(C₇) transfer with brachial plexus root avulsion and the effect on the contralateral limbs after operation. Methods: A total of 83 patients with injory of brachial plexus avulsion were underwent contralateral C₇ transfer from September, 2004 to December, 2014. Among them, contralateral C₇ were transferred to the median nerve in 33 cases, to radial nerve in 14 cases, to musculocutaneous nerve in 22 cases, and simultaneous transfer to median and musculocutaneous nerve in 14 cases. The followed-up contents included recovery of muscular strength and sensory innervated by the recipient nerve, bilateral limbs synergistic activity and the effect on the contralateral limbs after the surgery. Results: All 83 cases were successfully followed-up for 3.2-6.2 years (average, 4.5 years), and found no significant effect on function of the contralateral limbs. In the group of C₇ transferred to the median nerve group, 10 cases had muscle strength recovered≥M₃, and 26 cases had sensory recovered≥S₃; In the group of C₇ transferred to the radial nerve, 6 cases had muscle strength recovered≥M₃, and 9 cases had sensory recovered ≥S₃; In the group of C₇ transferred to the musculocutaneous nerve, 12 cases had muscle strength recovered≥M₃, and 17 cases had sensory recovered≥S₃. In the group of C₇ simultaneous transferred to the median and musculocutaneous nerve, 6 cases had muscle strength of wrist and digital flexion recovered≥M₃, 5 cases had elbow flexion recovered≥M₃, 9 and 10 cases had sensory of innervated area and lateral forearm region recovered≥S₃, respectively. Conclusion Contralateral C₇ transfer is an ideal procedure for the treatment of total brachial plexus root avulsion with definitive clinical outcomes.

Objective To study the clinical efficacy of two-stage bone transport combined with induced membrane technique in the treatment of post-traumatic tibial osteomyelitis.Methods A retrospective study was conducted of the 14 patients with post-traumatic tibial osteomyelitis who had been treated by two-stage bone transport combined with induced membrane technique from February 2009 to August 2015.They were 12 males and 2 females,aged from 26 to 59 years (average,40.4 years).By the Cierny & Mader classification,they all belonged to type Ⅳ.Their disease course ranged from 6 to 30 months,averaging 15.3 months.The number of their previous operations ranged from 2 to 6 times,averaging 3.3 times.In the first stage of operation,tibial extended external fixation was installed according to preoperative design,followed by early debridement and extensive removal of osteomyelitis lesion.The length of bone defect after resection of osteomyelitis lesion was from 4 to 18 cm (average,8.5 cm).10％ vancomycin-contained polymethyl methacrylate (PMMA) bone cement was molded and placed in the bone defects.In the second stage of operation,the PMMA-induced membrane was opened to remove the PMMA bone cement,the bone marrow was drilled through and the induced membrane was then sutured up.Tibial osteotomy was conducted at an appropriate part.The average interval between the first and second stages was from 4 to 7 weeks (average,4.8 weeks).The clinical efficacy was evaluated according to the Paley criteria at final follow-ups.Complications were recorded.Results All the patients were followed up for 15 to 50 months (17.5 months).No relapse of osteomyelitis occurred.The total treatment time ranged from 11 to 19 months,with an average of 14.7 months.The total time for external fixation was from 10 to 18 months,averaging 13.2 months.The external fixator indexes ranged from 31 to 75 day/cm,with an average of 48.5 day/cm.According to Paley criteria at final follow-ups,the clinical efficacy was excellent in 11 and good in 3 cases.Pin tract infection occurred in 9 patients,mal-alignment after removal of external fixation in 6,and malunion or nonunion at the docking site in 5.Conclusions Two-stage bone transport combined with induced membrane technique can cure thoroughly post-traumatic tibial osteomyelitis.Its operation procedures are simple but effective.

OBJECTIVETo review kidney transplantation in the center and analyze the risk factors affecting long-term allograft survival.

METHODSThirty-two relative variables were analyzed with SAS statistical software. Using Log-rank method, we investigated influence of these variables on short-and long-term survival of grafts. Kaplan-Meier analysis was used to estimate the 1-, 3-, 5-, 10-years graft survival rates and half-life. Proportional hazards regression analysis (Cox model) was used to assess and rank the relative risk of potential variables.

RESULTSThe 1-, 3-, 5-, 10-years graft survival rates were 83%, 75%, 66% and 48%. After excluding the patients died with functioning grafts, the 1-, 3-, 5-, 10 years grafts survival rate increased to 89%, 82%, 75% and 69%, respectively. The mean half-life was 8.78 +/- 0.14 and 14.09 +/- 0.20 years, respectively. By Log-rank analysis, factors affecting short- and long-term graft survival were identified as: renal function, duration of graft function became normal, cold-ischemia time, presence of acute rejection, delayed graft function, immunosuppressive regimen, complication, infection, anti-rejection therapy. Cox model multivariate analysis showed that there were 18 factors affecting graft survival.

CONCLUSIONSNew immunosuppressive agents not only significantly increase short-term graft survival, but also have the better long-term outcome tendency. Making assurance to get high quality donor organ and minimizing the death with graft function may be the most feasible way to prolong graft survival at present.

Adult ; Cadaver ; Female ; Graft Survival ; drug effects ; Humans ; Immunosuppressive Agents ; pharmacology ; Kidney Transplantation ; Male ; Multivariate Analysis ; Transplantation, Homologous

Objective:To perform trans-peritoneal laparoscopic adrenalectomy via a single incision in treatment of adrenal tumors,and to discuss its clinical outcome and safety.Methods:Single incision trans-peritoneal laparoscopic adrenalectomy was used in treatment of three patients with adrenal tumors.The incision was made 3 cm below the costal margin of anterior axillary line; three Tocars were placed in the cut.The instruments used included single port access,CUSA,Hem-o-lok,etc..Results:The three operations were all successful,and there were no conversion to open procedure or a need for extra Ttrocars.The operating time periods were 75,116,and 135 min,with a mean of (108.7?30.7)min.The perioperative blood losses were 10,20,and 30 ml,with a mean of (20?10)ml.The gastric canal and ureteral catheter were withdrew one day after operation,and the drainage tube was withdrew 3 d after operation.The mean postoperative hospital stay was 4 d.Conclusion:Single incision trans-peritoneal laparoscopic adrenalectomy has the advantage of little trauma,less blood loss,satisfactory safety,and prompt postoperative recovery,but is difficult to manage.