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.

ObjectiveTo evaluate term efficacy of focal cryoablation for localized single lesion prostate cancer and the protection of erectile function.MethodsFrom June 2008 to December 2009, 12 patients with localized single lesion prostate cancer were treated with focal cryoablation. All diagnoses were confirmed by transrectal ultrasound-guided prostatic biopsy. The mean Gleason score was ≤ 7; three cases had a Gleason score of 5, five cases a score of 6, and four cases a score of 7. The average preoperative value of PSA was 9.7 ng/ml ( 4.2 - 14.9 ng/ml ). The PSA level was obtained every three months for the first year and then every six months thereafter. Patients with a PSA nadir greater than 1.0 ng/ml or if the PSA nadir increased more than 2 ng/ml underwent repeat biopsy to exclude tumor recurrence.ResultsThe mean operative time was (82 ±26) minutes. The mean postoperative hospital stay was (5 ± 2) days. After catheter removal, all 12 cases had good continence and 10 had erectile function before surgery. Of the 10 patients, eight had erectile function after surgery. The follow - up ranged from 12 to 30 months ( mean 23 ).The lowest average serum PSA nadir was 2.2 ng/ml, nine patients had a minimum PSA nadir of less than 1.0 ng/ml and three had a minimum PSA nadir of more than 1.0 ng/ml. Four patients underwent repeat biopsy after abnormal PSA, and prostate cancer was detected in the contralateral gland in one patient.ConclusionsUltrasound-guided transperineal focal cryoablation is a safe and effective treatment with few complications for localized single lesion prostate cancer. Additional studies with longer follow-up are necessary for long-term efficacy.

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.

OBJECTIVETo study the stomach role in exogenous oxalate absorption.

METHODSThe kinetic variation of urinary oxalate excretion (mg/min) in 10 healthy adults and 8 patients who underwent total gastrectomy was investigated before and after an oral spinach oxalate load. The bioavailability of the oxalate load in the healthy adults was calculated and compared with that in the patients.

RESULTSThe oxalate content in the oral spinach load was 2567-2670 mg. The urinary oxalate excretion (mg/min) in the 10 healthy adults increased significantly 20 minutes after loading (this increase was compared against their basic oxalate excretion level of 0.0331 +/- 0.0203). Further observations after loading include: a first peak of oxalate excretion 40 minutes after loading; an oxalate excretion level double that of the basic level after 60 minutes (0.0732 +/- 0.0294) and a second peak appearing at 3 hours (P < 0.01). A "first peak" (0.063% +/- 0.062%) was not in any of the patients who underwent a total gastrectomy. Furthermore, a bioavailability of oxalate, which was 50% lower than that in the healthy subjects, appeared 60 minutes after loading (0.098% +/- 0.071%, P < 0.01).

CONCLUSIONSThe stomach is a powerful oxalate absorption organ under normal physiological conditions. Further investigation on the relationship between stomach dysfunction and urinary calcium oxalate formation is needed.

Absorption ; Adult ; Female ; Humans ; Male ; Middle Aged ; Oxalates ; pharmacokinetics ; Spinacia oleracea ; metabolism ; Stomach ; metabolism

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 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.