BACKGROUND: A middle cerebral artery occlusion and reperfusion(MCAO/R) model in rats with suture has been widely used in the researches of acute focal ischemic cerebral infarction, while the model in rabbits by the same method is relatively rare. Magnetic resonance diffusion weighted imaging (MR DWI) has been paid close attention recently for its sharp sensitivity of cerebral ischemia.OBJECTIVE: To establish rabbit models of MCAO/R by intraluminal thread, and study the characteristics of MR DWI after cerebral ischemia and reperfusion.DESIGN: Random controlled animal experiment.SETTING: Institute of Cerebrovascular Diseases, Affiliated Hospital of Qingdao University Medical College.MATERIALS: The experiment was accomplished at the Key Laboratory of Brain Diseases Prevention and Cure of Shandong Province from March to June in 2005. A total of 103 adult healthy New Zealand rabbits of either sex, 10-12 weeks old and 1.8-3.3 kg weight were provided by the Experimental Animal Center of Shandong Agricultural Academy (SCX20040013).They were bred at quiet, sanitary and dry conditions.METHODS: Animal groups: 103 rabbits were divided randomly into group A (n=53) and group B (n=50). The rabbits in group A were treated with suture of 0.51-0.55 mm as the diameter of thread, while group B was reassigned into B1 (0.46-0.50 mm), B2 (0.51-0.55 mm) and B3 (0.56-0.60 mm).The successful MCAO/R models in 57 cases were randomly divided into permanent ischemia group (n=30, ischemia 1, 3, 6, 12, 24 andl 48 hours, 5ones at each time point) and ischemic reperfusion group (n=27, reperfusion 0, 2 and 5 hours, 5 ones at each time point; reperfusion 11, 23 and 47hours, 4 ones at each time point). Another 10 rabbits receiving sham operations were regarded as contrasts for permanent ischemia group and ischemia reperfusion group, with 5 ones in each.MAIN OUTCOME MEASURES: The changes of hyperintensity area on DWI and apparent diffusion coefficient (ADC) were measured in permanent ischemia group and ischemic reperfusion group.RESULTS: The data of 57 successful model rabbits were involved in the result analysis.①The successful rate in group A (26 cases, 49.1%) was significantly lower than that in group B (31 cases, 62.0%).②In ischemia group:The hyperintensity area on DWI with declined ADC appeared at ischemia 1 hour. The hyperintensity areas on DWI at different times increased gradually from ischemia 1 hour and unchanged within 24 hours. The mean ADC at different times declined at first and then gradually increased.③In reperfusion group: Comparing with ischemia 1 hour, the hyperintensity area on DWI reduced while ADC increased at reperfusion 2 hours and 5 hours, and enlarged with ADC high at reperfusion 11 hours, then continued to enlarge with ADC reduced significantly at 23 hours and 47 hours.CONCLUSION: The diameter of thread tip and the inserting distance of thread are main factors for establishing successful MCAO/R models. The hyperintensity area on DWI and the decreasing ADC after acute cerebral ischemia can be improved by early reperfusion, but the secondary decreasing ADC may be induced by continuously reperfusion.

Objective To evaluate if the flexible ureteroscopy could treat stones located in lower calyx with the infundibulopelvic angle (IPA) less than 30°.Methods Thirty-six patients with inferior caliceal calculi on whom flexible ureteroscopic procedures were performed between November 2009 and June 2015 were reviewed.The mean age of the patients was 52.1 years (34-71),with the mean stone diameter of (1.5 ± 0.8) cm (1.2-2.6 cm).IPA was smaller than 30° in all 36 cases,which confirmed by CTU examination.IPA was less than 10° in 15 patients,between 11 ° and 20° in 13 patients and between 21 ° and 30° in 8 patients.Results The success rate was 63.9％ (23/36 patients) in patients with IPA smaller than 30° after first session of procedure,and the stone free rate reached 100％ after the second session of procedure.The mean operation duration was (95.5 ± 31.4) min(51-127 min).The mean hospital stay after operation was(4.1 ± 1.2)days (3-5 days).No major complications were recorded and no patients needed to convert to open surgery.Double J tube was removed after 4 weeks postoperatively.Patients were followed up for 4-12 months,during which ultrasound and CT scan were used for stone detection.Conclusions The small IPA (＜30°) negatively affected the SFR in the first session operation.However,its negative effect was solvable by using modern endoscopes.A complete stone clearance was achievable even in case of unfavorable anatomic conditions in experienced hand.

Objective To compare the therapeutic effect of retrograde ureteroscopic Ho:YAG laser lithotripsy (UHL) and percutaneous nephrolithotripsy (PCNL) in treating patients with renal calculi of ≤3 cm.Methods From Feb.2008 to Apr.2011,a total of 109 cases (117 renal calculi in total) treated by either PCNL (50) or UHL (67) were retrospectively analyzed.Operative time,stone free rate,postoperative hospital stay,hospitalization expenses and complications were compared in the 2 groups.Results The mean stone burden of the UHL group and PCNL group was 2.5 cm (1.5-3.0) cm and 2.4 cm (1.3-3.0) cm,respectively.There was no significant difference in two groups.In UHL group,operations were performed successfully in all 67 calculi,of whom,43 patients needed combination of flexible ureteroscopy to break the stone fragments falling into the renal calices.No serious complication was recorded except postoperative fever in 3 cases.In PCNL group,all patients had been successful operated.Postoperative fever occurred in 2 cases.Obvious intraoperative and postoperative haemorrhage appeared in 1 case,and cured by selective renal artery embolization.The operation time of the UHL group and PCNL group was (117.0±36.5) min and (90.0±18.3) min respectively,and had no significant difference in two groups (P＞0.05).The stone free rate of the UHL group and PCNL group was 94％ (63/67) and 92％ (46/50) respectively,and had no significant difference in two groups (P＞0.05).Hemoglobin decline of the UHL group and PCNL group was (0.3±0.1) g/L and (20.6±8.1) g/L,and had significant difference in two groups (P＜0.05).Postoperative hospital stay of the UHL group and PCNL group was (5.0±2.4) day and (7.0±3.7) day,and had statistical difference in two groups (P＜ 0.05).Hospitalization expenses of the UHL group and PCNL group was (15 477.0±754.3) RMB and (27 453.0± 1763.5) RMB,and had statistical difference in two groups (P＜0.05).Conclusions Although UHL and PCNL have similar curative effect in treating renal stones of less than 3 cm,UHL have the advantages of lcss trauma and complication,lower expenses and fewer hospital stay.In treating renal stones of smaller than 3 cm,especially in the contraindication of PCNL,UHL may be a better choice.

Objective To monitor the renal pelvic pressure and to investigate its clinical significance during retrograde flexible ureteroscopic lithotripsy (RFUL).Methods The data of renal pelvic pressure measured in 60 cases of RFUL with the mean irrigation pump speed and pressure of 30 ml/min and 30 mmHg were analyzed retrospectively.The influence factors of renal pelvic pressure and its correlation with postoperative fever were analyzed.Renal pelvic pressure was measured by baroceptor,which was connected to PHILIP-MP4 monitor IBP channel and ureteric catheter positioned in renal pelvis through a dual channel ureteral access sheath (UAS).The renal pelvic pressure data was collected and analyzed in every 2 seconds by computer.The 60 cases were divided into 3 groups according to their intra-pelvic pressure situations:normal pressure group(NP,IPPmax ≤30 mmHg),high pressure group(HP,IPPmax＞30 mmHg,but high pressure duration≤ 10 min),and backflow pressure group(BP,IPPmax＞30 mmHg and high pressure duration＞ 10 min).Results The baseline intra-pelvic pressure (IPP0) and max imum intra-pelvic pressure (IPPmax) were (13.2±5.6) mmHg and (95.6±2.3) mmHg respectively.IPP levels during the RFUL were significantly higher than the IPP0(P＜0.001).There were 32,17 and 11 cases in NP,HP and BP groups,respectively.There were 6 cases with fever higher than 38.5 ℃ (10％),in which there were 1 case in NP,1 case in HP group and 4 cases in BP group.The postoperative fever rate in NP,HP and BP group were 3％,6％ and 36％ respectively,which were significantly different between groups(P＜0.01).There were 12 cases with procalcitonin ＞0.1 ng/ml and 8 cases with procalcitonin ＞0.5 ng/ml,in which 2 cases in HP group and 6 cases in BP group.Conclusions RFUL would result in a temporal elevated intrapelvic pressure greater than 30 mmHg.Postoperative fever is relevant with renal perfusion pressure and perfusion time.It's necessary for the surgeons to adjust the perfusion pressure during operation.

The cloning and identification of frc gene from Oxalobacter formigenes in the intestines of Chinese people were conducted. The genomic DNA of Oxalobacter formigenes was extracted. frc gene fragment was amplified by polymerase chain reaction (PCR) and linked with pEGFP-C1. The recombinant plasmid was designated pEGFP-frc and was identified by restriction-enzyme digestion and sequencing. Human embryo kidney 293 cells were transfected with pEGFP-frc, then RT-PCR and Western blotting were performed to detect the expression of frc gene. The length of frc gene was found to be 1287 bp, and the homology of nucleotides and amino-acid residue with the sequence in GenBank was 95.88% and 99.07%. Bright green fluorescent light could be observed in 293 cells transfected with the pEGFP-frc. frc mRNA and fusion protein FCoAT-EGFP were detected in the cells. It is concluded that frc gene cloned from the Oxalobacter formigenes in the intestines of Chinese people can be expressed in eucaryotic 293 cells and keep its enzyme activity.

Objective To evaluate the clinical feasibility of radiofrequency ablation (RFA) for renal cell carcinoma in functional solitary kidney. Methods Five selected cases of primary renal cell carcinoma in a functional solitary kidney were retrospective analyzed. Three cases of left renal cell carcinoma and 2 cases of right renal cell carcinoma were diagnosed by B ultrasound and CT scanning. The maximum diameter of the tumors were 3.0- 6.0 cm. Three cases of which were T1a N0 M0 and the others Were T1bN0M0. All patients were followed up with enhanced-CT and contrast-enhanced ultrasonography. Results The mean time of the operation was 100.0+28.5 min, and the mean blood loss was 95.0±30.5 ml. No patients accepted blood transfusion post-operation. All patients after operation had fever (38.2±0. 7 ℃ ). All the lesions were completely ablated on contrast-enhanced ultrasonography. There was no statistic change of hemoglobin, serum creatinine (SCr), ECT-GFR and creatinine clearance after operation (P>0.05). All of the 5 cases survived. No recurrence were found by enhanced-CT and contrast-enhanced ultrasonography and SCr was retaining in the normal level during follow-ups (median, 11.5 months). Conclusion Radiofrequency ablation for renal cell carcinoma in a functional solitary kidney could be a treatment choice with a relatively low incidence of complications.

The cloning and identification of frc gene from Oxalobacter formigenes in the intestines of Chinese people were conducted. The genomic DNA of Oxalobacter formigenes was extracted. frc gene fragment was amplified by polymerase chain reaction (PCR) and linked with pEGFP-C1. The recombinant plasmid was designated pEGFP-frc and was identified by restriction-enzyme digestion and sequencing. Human embryo kidney 293 cells were transfected with pEGFP-frc, then RT-PCR and Western blotting were performed to detect the expression offrc gene. The length of frc gene was found to be 1287 bp, and the homology of nucleotides and amino-acid residue with the sequence in GenBank was 95.88% and 99.07%. Bright green fluorescent light could be observed in 293 cells transfected with the pEGFP-frc. frc mRNA and fusion protein FCoAT-EGFP were detected in the cells. It is concluded that frc gene cloned from the Oxalobacterformigenes in the intestines of Chinese people can be expressed in eucaryotic 293 cells and keep its enzyme activity.

Objective:To investigate the promotion effect of 4,5,6,7-tetrabromobenzotriazole (TBB)on the apoptosis of human colon cancer SW480 cells,and to explore its possible mechanism.Methods:The human colon cancer SW480 cells at logarithmic growth phase were divided into control group (0 μmol·L-1 TBB)and experiment group (1,3,10,30,and 100 μmol·L-1 TBB).The viability of cells was measured by MTT assay;the apoptotic rate of the SW480 cells and the level of intracellular reactive oxygen species (ROS)were analyzed by Annexin Ⅴ-FITC/PI double staining and flow cytometry.The expression levels of anti-apoptotic proteins p-Akt and Bcl-2,and pro-apoptotic proteins Bad, pro-caspase-9 and cleaved-caspase-3 were detected by Western blotting method. Results:The MTT results showed that the viabilities of SW480 cells in experiment group were decreased in a dose-dependent manner,which were lower than that in control group (P < 0.05).The Annexin Ⅴ-FITC/PI double staining and flow cytometry results showed that the apoptotic rates of SW480 cells in experiment group (3,6,12, and 24 h)were significantly higher than that in control group (0 h)(P <0.05).The flow cytometry results showed the levels of ROS in SW480 cells after treated with TBB for 3,6,12 and 24 h were higher than that in 0 h group (P <0.05).The Western blotting results showed that the expression levels of anti-apoptotic proteins p-Akt and Bcl-2 in SW480 cells in experiment group (3,6,12,and 24 h)were decreased obviously,whereas the expression levels of the pro-apoptotic proteins Bad and cleaved-caspase-3 were increased and the expression level of pro-caspase-9 was decreased compared with those in control group (0 h) (P < 0.05 ).Conclusion: TBB could inhibit the cell proliferation and induce the apoptosis of human colon cancer SW480 cells,and its mechanism may be related to the inhibition of the activity of Akt and the promotion of the level of intracellular ROS.

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.