[Comparison of application effects of colonoscopy, fecal immunochemical test and a novel risk-adapted screening approach in colorectal cancer screening in Xuzhou population].

Objective: To compare the application effect of the colonoscopy, fecal immunochemical test (FIT) and novel risk-adapted screening approach in colorectal cancer screening in Xuzhou population. Methods: From May 2018 to April 2019, 4 280 subjects aged 50-74 were recruited from Gulou district, Yunlong district and Quanshan district of Xuzhou. They were randomly assigned to the colonoscopy group (n=863), FIT group (n=1 723) and novel risk-adapted screening approach group (n=1 694) according to the ratio of 1∶2∶2. For the novel risk-adapted screening approach group, after the risk assessment, high-risk subjects were invited to undergo colonoscopy and low-risk subjects were invited to undergo FIT examination. All FIT positive subjects were invited to undergo colonoscopy. Colonoscopy participation rate [(the number of colonoscopies completed/the number of colonoscopies invited to participate)×100%], detection rate of colorectal lesions [(the number of diagnosed patients/the number of colonoscopies completed)×100%], colonoscopy resource load (the number of colonoscopies completed/the number of diagnosed advanced tumors) and FIT resource load in each group were calculated and compared. Results: The age of all subjects was (61±6) years old, including 1 816 males (42.43%). There was no statistically significant difference in the socio-demographic characteristics of the subjects in different screening groups. The colonoscopy participation rate was 22.60% (195/863) in the colonoscopy group, 57.04% (77/135) in the FIT group, and 33.94% (149/439) in the novel risk-adapted screening approach group, respectively. The colonoscopy participation rate was higher in the FIT group than in the colonoscopy group and the novel risk-adapted screening approach group (P<0.001). The colonoscopy participation rate of novel risk-adapted screening group was significantly higher than the colonoscopy group (P<0.001). The detection rates of advanced tumors were 6.67% (13/195), 9.09% (7/77) and 8.72% (13/149), respectively, and the difference was not statistically significant (P>0.05). The colonoscopy resource load (95%CI) was 15 (13-17) in the colonoscopy group, 11 (9-14) in the FIT group and 11 (10-13) in the novel risk-adapted screening approach group, respectively. Among them, the colonoscopy resource load of high-risk individuals in the novel risk-adapted screening approach group was 12 (9-15). FIT resource loads (95%CI) were 207 (196-218) and 88 (83-94) in the FIT group and the novel risk-adapted screening approach group. Conclusion: The combined application of risk-adapted screening approach and FIT may have a good application effect in colorectal cancer screening.

[Controversy and strategy of partial versus total adrenalectomy in the treatment of primary aldosteronism].

Primary aldosteronism (PA) patients diagnosed with aldosterone-producing adenoma and unilateral adrenal hyperplasia are preferred for surgical treatment.The aim of the operation is to remove the hypersecretory aldosterone tissue of adrenal and to improve the prognosis.Current studies have shown that both partial adrenalectomy and total adrenalectomy can achieve clinical cure for PA, but the choice of surgical methods is still controversial. This paper discusses the efficacy and safety, postoperative changes in cortisol level, postoperative recurrence and surgical techniques of different surgical methods in the treatment of different PA subtypes, so as to find the best surgical methods for different PA subtypes and to maximize the benefits for patients.

[Practice of laparoscopic anatomical adrenalectomy].

On the basis of existing laparoscopic transabdominal adrenalectomy, this article described a safe and reliable surgical method for the treatment of small adrenal lesions-laparoscopic anatomical adrenalectomy (LAA), and retrospectively analyzed the clinical data of 74 patients who had undergone LAA. All patients had no signs of recurrence on imaging. LAA has high safety and feasibility, clear intraoperative anatomical layers, good spatial operability, and low postoperative complications. LAA provides a more reliable option for the treatment of small adrenal diseases.

[Effect of total prostate volume and transitional zone volume on the curative effect of transurethral resection of prostate in benign prostatic hyperplasia].

Objective: To compare the clinical effect of different total prostate volume (TPV) and different transitional zone volume (TZV) on benign prostatic hyperplasia (BPH) treated with transurethral resection of prostate(TURP). Methods: Clinical data of 210 patients with BPH admitted to Guizhou provincial people's hospital from June 2016 to August 2018 were retrospectively collected and analyzed. All patients underwent transrectal ultrasonography, and they were divided into three groups according to TPV: 70 patients in group A:TPV<40mL, 98 patients in group B: 40 ml≤TPV<80 ml, and 42 patients in group C:TPV≥80 ml. Meanwhile, three groups were divided according to TZV: 88 patients in group a: TZV<20ml, 67 patients in group b: 20 ml≤TZV<40 ml, and 55 patients in group c:TZV≥40 ml. All of the patients with TURP were followed up for 6 months after surgery, and the data of international prostate symptom score (IPSS), storage symptoms IPSS (IPSS-S), voiding symptoms IPSS (IPSS-V), Quality of Life (QoL) index, and maximum urinary flow rate (Qmax) were collected before and after surgery. Finally, the effect of TPV and TZV on TURP was analyzed respectively by analysis of variance. Results: There were no statistically significant differences in preoperative IPSS, IPSS-V, IPSS-S, QoL and Qmax among the three groups of patients grouped by TPV (P>0.05), but the age of patients in group C(73.5±6.5) was significantly higher than that in group A (69.3±7.6) and group B (70.9±7.3) (P=0.015). Postoperative IPSS, IPSS-V, IPSS-S, QoL and Qmax of patients in groups A, B and C also showed no significant difference (P>0.05). There were no statistically significant differences in preoperative IPSS, IPSS-V, IPSS-S, QoL, and Qmax among the three groups of patients grouped according to TZV (P>0.05), while the age of patients in group a (69.2±7.6) was significantly lower than that of patients in group b (72.1±7.2) and group c (72.5±6.7) (P=0.017). There were statistically significant differences in IPSS (P=0.010), IPSS-V (P=0.037), IPSS-S (P=0.022), QoL (P=0.038) and Qmax (P=0.037) among the groups a, b, and c after surgery. Moreover, IPSS, IPSS-V, IPSS-S and QoL were negatively correlated with TZV, while Qmax was positively correlated with TZV. Postoperative IPSS, IPSS-V, IPSS-S, QoL and Qmax were significantly different from those before surgery in groups A, B, C and groups a, b, c (P<0.001). Conclusion: TPV and TZV may not be significantly correlated with BPH symptoms, but may be correlated with age. TURP is an effective treatment for patients with different TPV and TZV. There is no significant statistical difference in the surgical efficacy among patients with different TPV, but patients with larger TZV tended to have better outcome. TZV may be better than TPV in predicting the postoperative efficacy.

[The potential application of a three-dimensional visualization technique in adrenal vein sampling].

Objective: To explore the potential application of a three-dimensional visualization technique in adrenal vein sampling (AVS). Methods: The clinical data were retrospectively analyzed, which included 76 patients with primary aldosteronism (PA) who have undergone AVS in Guizhou Provincial People's Hospital from December 2017 to May 2020. All cases were examined by adrenal thin-section enhanced CT and blood was drawn by bilateral AVS. Among them, the adrenal vein blood of 46 cases was sampled with the help of three-dimensional (3D) visualization processing of CT data, while that of 30 cases was without 3D visualization processing. The rate of the catheter in place, the successful rate of AVS, the time of blood collection, the dosage of the contrast agent, and surgical complications were compared between the two groups. Results: There were 76 cases included, while 38 were male and 38 were female. The average age was 45 (25-57) years. Compared with the patients without the aid of 3D visualization, the success rate of right AVS of the patients with the aid of 3D visualization technology increased from 43% to 78% (P<0.05). The success rate of adrenal vein blood collection increased from 53% to 83%. The dose of contrast agent decreased [the M(Q1,Q3) were78 (59, 89) ml vs 28 (16, 51) ml, P<0.05], and the time of blood sampling from the right adrenal vein approximately decreased [the M（Q1，Q3） were 70 (66, 88) min vs 44 (22, 61) min, P<0.05]. Compared with the case without the aid of 3D visualization, the left adrenal vein catheterization rate of patients in the 3D visualization group increased from 97% to 98%, the success rate of adrenal vein blood collection increased from 97% to 98%, and the differences of the time of blood sampling and the dosage of the contrast were not statistically significant between the two groups. Among all the cases experienced bilateral AVS, only one patient without 3D reconstruction had contrast extravasation, and the others had no obvious complications. Conclusions: Before AVS, 3D visualization processing of adrenal vein from CT data is capable of increasing the success rate of blood sampling from the right adrenal vein, as well as reducing the dosage of contrast agent and the time of adrenal vein blood sampling. Therefore, it has a potential clinical value of the application.

[Effect of transition zone index on the outcome after transurethral resection of prostate].

Objective: To investgate the effect of properative transitional zone index (TZI) on the outcome of transurethral resection of prostate (TURP). Methods: A retrospective analysis was performed on 206 patients with TURP who were admitted to our hospital from January 2016 to September 2018. All patients underwent transrectal ultrasound (TRUS) to determine the total prostate volume (TPV) and the transition zone volume (TZV). Patients were divided into two groups according to TZI (TZV/TPV) (group A: TZI<0.5, group B: TZI≥0.5). We collected data 6-months after surgery including international prostate symptom score (IPSS), quality of life score (QOL), maximum flow rate (Qmax), and postvoiding residue (PVR) to compare the difference of the postoperative outcome of two groups, while the IPSS was subdivided into voiding (IPSS-v) and storage(IPSS-s) symptoms, and the changes of IPSS, IPSS-v, IPSS-s, QOL, Qmax and PVR before and after surgery were analyzed. The treatment efficacy was determined as the changes of IPSS (post/preoperative IPSS: %IPSS), QOL (preoperative QOL-postoperative QOL: ΔQOL) and Qmax(preoperative Qmax-postoperative Qmax: ΔQmax). Pearson linear correlation analysis was employed to estimate the correlation of TZI and %IPSS, ΔQOL and ΔQmax, respectively. Results: A total of 126 patients were in Group A, and 80 patients were in group B. 1. The preoperative clinical data of the two groups were compared. There were no significant differences regarding age, IPSS, IPSS-v, IPSS-s, QOL, Qmax, and PVR between two groups (all P>0.05). However, the TPV of patients in group B (74.57±29.25) ml was significantly larger than that in group A (46.25±24.56) ml, P<0.001. While the postoperative follow-up outcomes of the two groups were compared, we found that IPSS-s (P=0.079), QOL (P=0.710), and PVR (P=0.651) were not statistically different between the two groups, but the postoperative IPSS, IPSS-v, and Qmax (8.50±5.75 vs 6.38±4.36, 4.03±3.75 vs 2.63±2.5, and (16.54±4.43) ml/s vs (18.94±4.84) ml/s, all P<0.05) were significantly different between the two groups. 2. Postoperative IPSS, IPSS-v, IPSS-s, QOL, Qmax and PVR were significantly different from those before surgery in two groups, respectively. 3. Pearson linear correlation analysis showed a significant positive correlation between TZI and ΔQmax (r=0.32, P<0.01), a weaker negative correlation between TZI and %IPSS (r=-0.22, P<0.01), and no correlation between TZI and ΔQOL (r=0.08, P=0.238). Conclusion: There may be a correlation between the outcome of TURP and TZI, and the outcome of TURP may be better in patients with TZI ≥ 0.5.

[Drugs endocrine therapy improves lower urinary tract symptoms in patients with prostate cancer and analysis of related factors].

Objective: To study the improvement of lower urinary tract symptoms and related influencing factors in patients with prostate cancer complicated with lower urinary tract symptoms after drugs endocrine therapy. Methods: The clinical data of 80 patients with prostate cancer with lower urinary tract symptoms and receiving drug endocrine therapy in Guizhou Provincial People's Hospital from March 2015 to June 2019 were analyzed retrospectively. All patients were followed up, and the mode of administration of the endocrine therapy, the time of administration, and the international prostate symptom score (IPSS2) and quality of life score (QOL2) after treatment were recorded. The improvement of IPSS score and quality of life score after endocrine therapy was observed. The correlation between variance and age, the prostate volume, PSA, tumor stage, Gleason score, symptom severity, medication mode, and medication time were analyzed by variance, chi-square test and paired sample t test. Results: The differences of QOL and IPSS before and after therapy were statistically significant (P<0.05). Chi-square test found that the effective rate of drug endocrine therapy was not related to age (χ2=0.800; P=0.371), tumor stage (χ2=0.094; P=0.759), PSA (χ2=0.651; P=0.420), prostate volume (χ2=0.216; P=0.642), Gleason score (χ2=0.157; P=0.692), symptom severity (χ2=0.457; P=0.499), medication mode (χ2=2.910; P=0.233), and medication time (χ2=4.159; P=0.385). Analysis of variance found that prostate volume and the severity of symptoms was significantly associated with improvement in lower urinary tract symptoms. Conclusions: Drug endocrine therapy can effectively improve the lower urinary tract symptoms of patients with prostate cancer, and the more severe the symptoms and prostate volumeof the patients, the more obvious the improvement of lower urinary tract symptoms. The age, tumor stage, PSA, Gleason score, time and manner of administration were not significantly correlated with improvement in lower urinary tract symptoms.

[The pathological characters of urothelial barrier in prostatic ducts of elderly SD rats].

Objective: To investigate the pathological characters of urothelial barrier in prostatic ducts of elderly male SD rats. Methods: Ten elderly male SD rats were anesthetized with chloral hydrate and then took the bladder, prostatic urethra and prostate tissues for serial pathological sections and HE staining. Immunohistochemical method was used to detect the expression of the Cytokeratin 7 (CK7), Cytokeratin 20 (CK20), Uroplakin Ⅲ (UP Ⅲ), Zonula occludens-1 (ZO-1) and Occludin in issues, and to make densitometric analysis (IA) on immunohistochemical results of each antibody. Results: HE staining observed that urothelial umbrella cells exist in the bladder, prostatic urethra and proximal prostatic duct. Immunohistochemical method showed the CK7, UPⅢ, ZO-1 and Occludin were positive in bladder, prostatic urethra and prostatic duct, while CK20 was negative. CK7 and UPⅢ were positive in the proximal prostatic duct, and negative in the bottom of the ductal acinar lumen; ZO-1 and Occludin were positive in prostatic duct. The IOD/Area values of CK7 and UP Ⅲ, in bladder, prostatic urethra, proximal prostatic duct, the middle of prostatic duct and bottom of the ductal acinar lumen, were 0.16, 0.13, 0.06, 0.05, 0.00 and 0.17, 0.16, 0.08, 0.05, 0.00(P<0.05,respectively). The expression of ZO-1 and Occludin in bladder (0.14 and 0.13) were higher than those in other tissues (0.11-0.12 and 0.09-0.10, P<0.05); the expression of ZO-1 and Occludin, which in proximal prostatic duct to the middle of prostatic duct and bottom of the ductal acinar lumen, had no significant difference (0.12-0.11 and 0.09-0.09, P>0.05). The IA values of CK20 were extremely low (0.00-0.01, P>0.05). Conclusion: Urothelial barriers partially exist in the prostatic ducts of elderly male SD rats, and with the prostatic ducts gradually extending to the bottom of acinar lumen, the urothelial barriers disappear. The results lay a foundation for further study on the repair of urinary epithelial barrier after prostatectomy.

[Comparative observation of thulium laser resection of the prostate-tangerine technique and transurethral resection of the prostate for the treatment of benign prostatic hyperplasia of various sizes].

Objective: To compare the efficiency and safety of thulium laser resection of the prostate-tangerine technique (TmLRP-TT) and transurethral resection of the prostate (TURP) for the treatment of benign prostatic hyperplasia (BPH) of various sizes. Methods: Clinical data of 249 BPH patients received TmLRP-TT or TURP were retrospectively collected. Patients were divided into small prostate group [prostate volume (PV)<40 ml], medium prostate group (40 ml≤PV<80 ml) and large prostate group (PV ≥ 80 ml) based on transrectal ultrasound (TRUS) results. Age, PV, Prostate-specific antigen (PSA), International prostate symptom score (IPSS), Quality of life (QoL), maximum of flow rate (Q(max)) and post-void residual urine (PVR) of patients received TmLRP-TT or TURP in each group were analyzed, as well as the perioperative data including operation time, hemoglobin and serum sodium level, transfusion, postoperative length of indwelling catheter and postoperative hospital stay. Furthermore, the postoperative complication rates of patients received the two operative methods in each group up to follow-up of 6 months were compared. Results: As for baseline indicators, there were no significant differences regarding age, prostate volume, PSA, IPSS, QoL, Qmax and PVR of patients received TmLRP-TT or TURP in each group (all P>0.05). In the small prostate group, there were no significant differences with operation time, hemoglobin and serum sodium level, transfusion, postoperative length of indwelling catheter and postoperative hospital stay received TmLRP-TT or TURP (all P>0.05). For the medium prostate group, patients received TmLRP-TT underwent longer operation time [(67.4±15.1) vs (57.5±11.5) min, P<0.001], but shorter length of indwelling catheter [(1.5±0.6) vs (3.1±0.9) d, P<0.001] and postoperative hospital stay [(3.5±0.9) vs (5.6±1.0) d, P<0.001], and there were no significant differences regarding transfusion rate (3/73 vs 1/78, P=0.280), hemoglobin [(9.8±9.0) vs (12.2±9.6) g/L, P=0.107] and serum sodium decrease [(2.07±3.65) vs (2.97±3.35) mmol/L, P=0.373]. In the large prostate group, patients received TmLRP-TT also underwent longer operation time [(86.5±14.3) vs (76.7±14.6) min, P=0.022], but less hemoglobin [(11.3±13.8) vs (23.3±15.0) g/L, P=0.006] and serum sodium decrease [(2.41±2.67) vs (4.00±6.22) mmol/L, P=0.042], lower transfusion rate (5/27 vs 0/24, P=0.026), and shorter length of indwelling catheter [(1.8±0.7) vs (4.3±1.5) d, P<0.001] as well as postoperative hospital stay [(3.7±1.1) vs (6.1±1.7) d, P<0.001]. Less overall complications were encountered in the medium (38/73 vs 24/78, P=0.008) and large (26/27 vs 10/24, P<0.001) prostate group who received TmLRP-TT, which was not seen in the small prostate group (P=0.589). Conclusions: TmLRP-TT and TURP are similarly efficient for the treatment of BPH of various sizes. For BPH patients with medium and large prostate, TmLRP-TT demonstrated significant advantages in reducing the overall complications, although the operation time was slightly longer.

[Efficacy and safety of pegylated interferon α-2b injection (Y shape, 40 kD) in treatment of patients with genotype 1/6 chronic hepatitis C].

Objective: To investigate the efficacy and safety of the new investigational drug pegylated interferon α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 µg/week) combined with ribavirin in the treatment of patients with genotype 1/6 chronic hepatitis C (CHC), with standard-dose Peg-IFN-α-2a combined with ribavirin as a positive control. Methods: A multicenter, randomized, open-label, and positive-controlled phase III clinical trial was performed. Eligible patients with genotype 1/6 CHC were screened out and randomly divided into Peg-IFN-α-2b(Y shape, 40kD) group and Peg-IFN-α-2a group at a ratio of 2:1. The patients in both groups were given oral ribavirin for 48 weeks in addition and then followed up for 24 weeks after drug withdrawal. Abbott Real Time HCV Genotype II was used to determine HCV genotype, and Cobas TaqMan quantitative real-time PCR was used to measure HCV RNA level at 0, 4, 12, 24, 48, and 72 weeks. Adverse events were recorded in detail. The primary efficacy endpoint was sustained virological response (SVR), and a non-inferiority test was also performed. Results: A total of 561 patients with genotype 1/6 CHC were enrolled, among whom 529 received treatment; 90.9% of these patients had genotype 1 CHC. The data of the full analysis set showed that SVR rate was 69.80% (95% CI 65.00%-74.60%) in the trial group and 74.16% (95% CI 67.73%-80.59%) in the control group (P = 0.297 0). The data of the per protocol set (PPS) showed that SVR rate was 80.63% (95% CI 76.04%-85.23%) in the trial group and 81.33% (95% CI 75.10%-87.57%) in the control group (P = 0.849 8), and the 95% CI of rate difference conformed to the non-inferiority standard. The analysis of the PPS population showed that of all subjects, 47.9% achieved rapid virologic response, with a positive predictive value of 93.8%. The incidence rate of adverse events was 96.30% in the trial group and 94.94% in the control group, and the incidence rate of serious adverse events was 5.13% in the trail group and 5.06% in the control group. Conclusion: In the regimen of Peg-IFN-α combined with ribavirin for the treatment of genotype 1/6 CHC, the new investigational drug Peg-IFN-α-2b(Y shape, 40 kD) has comparable clinical effect and safety to the control drug Peg-IFN-α-2a.

[Clinical effect and safety of pegylated interferon-α-2b injection (Y shape, 40 kD) in treatment of HBeAg-positive chronic hepatitis B patients].

Objective: To investigate the clinical effect and safety of long-acting pegylated interferon-α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 µg/week) in the treatment of HBeAg-positive chronic hepatitis B (CHB) patients, with standard-dose Peg-IFN-α-2a as positive control. Methods: This study was a multicenter, randomized, open-label, and positive-controlled phase III clinical trial. Eligible HBeAg-positive CHB patients were screened out and randomized to Peg-IFN-α-2b (Y shape, 40 kD) trial group and Peg-IFN-α-2a control group at a ratio of 2:1. The course of treatment was 48 weeks and the patients were followed up for 24 weeks after drug withdrawal. Plasma samples were collected at screening, baseline, and 12, 24, 36, 48, 60, and 72 weeks for centralized detection. COBAS® Ampliprep/COBAS® TaqMan® HBV Test was used to measure HBV DNA level by quantitative real-time PCR. Electrochemiluminescence immunoassay with Elecsys kit was used to measure HBV markers (HBsAg, anti-HBs, HBeAg, anti-HBe). Adverse events were recorded in detail. The primary outcome measure was HBeAg seroconversion rate after the 24-week follow-up, and non-inferiority was also tested. The difference in HBeAg seroconversion rate after treatment between the trial group and the control group and two-sided confidence interval (CI) were calculated, and non-inferiority was demonstrated if the lower limit of 95% CI was > -10%. The t-test, chi-square test, or rank sum test was used according to the types and features of data. Results: A total of 855 HBeAg-positive CHB patients were enrolled and 820 of them received treatment (538 in the trial group and 282 in the control group). The data of the full analysis set showed that HBeAg seroconversion rate at week 72 was 27.32% in the trial group and 22.70% in the control group with a rate difference of 4.63% (95% CI -1.54% to 10.80%, P = 0.1493). The data of the per-protocol set showed that HBeAg seroconversion rate at week 72 was 30.75% in the trial group and 27.14% in the control group with a rate difference of 3.61% (95% CI -3.87% to 11.09%, P = 0.3436). 95% CI met the non-inferiority criteria, and the trial group was non-inferior to the control group. The two groups had similar incidence rates of adverse events, serious adverse events, and common adverse events. Conclusion: In Peg-IFN-α regimen for HBeAg-positive CHB patients, the new drug Peg-IFN-α-2b (Y shape, 40 kD) has comparable effect and safety to the control drug Peg-IFN-α-2a.

The urogenital-hypogastric sheath: an anatomical observation on the relationship between the inferomedial extension of renal fascia and the hypogastric nerves.

PURPOSE: The study aimed to perform an anatomical observation on the inferomedial extension of the renal fascia (RF) to the pelvis and explore its relationship with the hypogastric nerves (HGNs). METHODS: Gross anatomy was performed on 12 formalin-fixed and 12 fresh cadavers. Sectional anatomy was performed on four formalin-fixed cadavers. RESULTS: Different from the traditional concept, both the anterior and posterior RF included the outer and inner layer with different inferomedial extensions. The multiple layers of RF extended downward to form a sandwich-like and compound fascia sheath with potential and expandable spaces which was named as "the urogenital-hypogastric sheath." Below the level of the origin of the inferior mesenteric artery, the bilateral urogenital-hypogastric sheath communicated with the counterpart in front of the great vessels in the midline and the superior hypogastric plexus ran into the urogenital-hypogastric sheath which carried the HGNs, ureters, and genital vessels downward to their terminations in the pelvis. In the retrorectal space, the urogenital-hypogastric sheath surrounded the fascia propria of the rectum posterolaterally as a layer of coat containing HGNs. CONCLUSION: The multiple layers of RF with different extensions are the anatomical basis of the formation of the urogenital-hypogastric sheath. As a special fascial structure in the retroperitoneal space and the pelvis, emphasis on its formation and morphology may be helpful for not only unifying the controversies about the relationship between the pelvic fascia and HGNs but also improving the intraoperative preservation of the HGNs by dissecting in the correct surgical plane.

Inhibition of connective tissue growth factor by small interfering RNA prevents renal fibrosis in rats undergoing chronic allograft nephropathy.

AIM: Connective tissue growth factor (CTGF) is a highly profibrogenic molecule implicated in renal fibrogenesis. Small interfering RNA (siRNA) is an effective tool to silence gene expression. This study determined whether caudal vein injection of siRNA targeting CTGF inhibited its expression in rat kidneys in vivo, and furthermore whether it protected the kidney from renal fibrosis in chronic allograft nephropathy (CAN). METHODS: Male inbred Fischer (F344, RT1(lv1)) rat renal grafts were orthotopically transplanted into Lewis (LEW, RT1(1)) rats following the procedure of Kamada with our modification. At 6 weeks, recipients were divided into siRNA, normal saline (NS), and control siRNA groups, using daily siRNA-targeting CTGF (0.1 mg/kg), or NS, or a control siRNA via caudal vein injection for 14 days. At 4, 6, and 8 weeks, we observed the pathologic changes, expression of CTGF, E-cadherin, collagen I and IV, and anti-smooth muscle actin (alpha-SMA). RESULTS: Serum creatinine level, Banff score, and the expression of CTGF were significantly lower among the siRNA than the NS or the control siRNA groups at 8 weeks (P < .05). The expressions of collagen I and IV, and alpha-SMA were also significantly downregulated and E-cadherin was lost in the siRNA versus the NS and control siRNA groups at 8 weeks. CONCLUSIONS: This study showed that delivery of CTGF siRNA via the caudal vein significantly inhibited expression of CTGF in rat kidneys, effectively preventing fibrosis in CAN. The results suggest that siRNA-targeting of CTGF has the potential to be a novel strategy for amelioration of CAN.

C4d deposition is associated with chronic allograft nephropathy in rats and could be influenced by immunosuppressants.

AIMS: Deposition of C4d in peritubular capillaries (PTC) has been considered to be a marker of humoral immunity in renal transplant. This study is to investigate C4d deposition in rat renal allografts undergoing CAN and the effects of immunosuppressants on it. METHODS: Fisher 344 rat renal grafts were orthotopically transplanted into Lewis rats following the procedure of Kamada with our modification. All the recipients were given CsA 10 mg/kg(-1).d(-1) x 10 d and then divided into 5 groups (each n = 9); (1) Vehicle: vehicle orally, (2) CsA: 6 mg/kg(-1).d(-1), (3) RAPA: 0.8 mg/kg(-1).d(-1), (4) FK 506: 0.15 mg/kg(-1).d(-1), (5) MMF: 20 mg/ kg(-1).d(-1). At 4 weeks, 8 weeks, 12 weeks, the rats were sacrificed, renal allografts were harvested and sera were collected. The deposition of C4d was detected by immunofluorescence and analyzed by Integrated Optical Density (IOD). The pathological changes were accessed according to the Banff 97 criteria. RESULTS: C4d deposition in PTC was found in all the allografts at 4 weeks, while there was no obvious manifestations of CAN in all the groups; the differences of Banff Score between all groups were not significant (P > .05). The values of IOD in RAPA and MMF group were lower than those in other 3 groups (P = .002, .006). The differences between RAPA and MMF, and between other 3 groups were not significant (P > .05). The intensity of C4d increased along with the progression of CAN, the heaviest C4d deposits in PTC were found at 12 weeks, and meanwhile the severest CAN was found. Comparing with Vehicle group, CsA and FK 506 had no effect on C4d deposition (P > .05), however, MMF and RAPA obviously decreased the C4d deposition (P = .000). The intensity of C4d deposition had a significant correlation with the severity of CAN (r = 0.894, P = .000). CONCLUSIONS: Our study suggests that the deposition of C4d in allografts appears earlier than pathological changes of CAN and has a correlation with the progression of CAN. MMF and RAPA can attenuate CAN by inhibiting humoral immunity. In contrast, CsA and FK 506 have no effect on humoral immunity.

C4d deposition is correlated with the level of antivimentin antibody in rat kidneys undergoing chronic allograft nephropathy.

AIMS: Antivimentin antibody is often produced as an autoantibody after transplantation. C4d deposition, a marker of humoral immunity during transplantation, is believed to reflect alloantibodies. This study investigated the relationship between C4d deposition and humoral immunity to vimentin among rat kidneys undergoing chronic allograft nephropathy (CAN). METHODS: Fisher 344 rat renal grafts were orthotopically transplanted into Lewis rats following the procedure of Kamada with our modification. All recipients were administered cyclosporine (CsA) (10 mg/kg(-1).d(-1) x 10 d) before being divided into 3 groups of oral treatments: (1) vehicle, (2) CsA (6 mg/kg(-1).d(-1)), and (3) mycophenolate mofetil (MMF; 20 mg/kg(-1).d(-1)). At 4, 8 and 12 weeks after transplantation, the rats were killed, the renal allografts harvested, and the sera collected. Serum creatinine (SCr) was measured and pathologic changes assessed according to the Banff 97 criteria. The antivimentin antibody was quantified by enzyme-linked immunosorbent assay. The deposition of C4d detected by immunofluorescence was analyzed by integrated optical density (IOD). RESULTS: Antivimentin antibody was observed in sera of all transplanted rats. The level of antivimentin antibody (IgGDeltaOD) increased gradually during the development of CAN from 4 weeks. Simultaneously, C4d deposition in peritubular capillaries also progressively strengthened. There was a strong positive correlation between the content of antivimentin antibody and C4d deposition (r = 0.892; P = .000). MMF simultaneously decreased antivimentin antibody formation and C4d deposition. In contrast, CsA had no significant effect. CONCLUSIONS: We demonstrated the production of antivimentin antibodies and the deposition of C4d during the development of CAN. There was a positive correlation between them. Whether humoral immunity to vimentin contributes to C4d deposition is not clear and further studies are needed to elucidate this issue.

Expressions of Angiopoietin-1, Angiopoietin-2, and Tie2 and their roles in rat renal allografts with chronic allograft nephropathy.

OBJECTIVE: Angiopoietin-1 (Ang1) and -2(Ang2) are 2 ligands for the endothelium-specific tyrosine kinase Tie2. Previous studies have shown that reciprocal regulation of Ang1, Ang2, and Tie2 plays an important role in chronic cardiac allograft vasculopathy. This study investigated the expressions of Ang1, Ang2, and Tie2 in rat renal allografts undergoing chronic allograft nephropathy (CAN). MATERIALS AND METHODS: Renal transplantations following the procedure of Kamada with our modification were orthotopically performed using Fisher (F344, RT1(1v1)) rats as both donors and recipients in the autograft group. Fisher and Lewis (LEW, RT1(1)) rats were used as donors and recipients in the allograft group, respectively, which was treated with cyclosporine (CsA; 10 mg/kg/d x 10 d). At 4w, 8w, and 12 weeks posttransplantation, serum creatinine (SCr) was measured and pathologic changes assessed according to the Banff 97 criteria. The mRNA (Deltact) and protein expressions of Ang1, Ang2, and Tie2 were localized by real-time fluorescence quantitative polymerase chain reaction (PCR) and by immunohistochemistry. RESULTS: The elevation in SCr and the pathologic changes in CAN were observed in all allografts at 8 and 12 weeks. The expressions of Ang1 and Ang2 were localized to epithelial cells and endothelium of the vascular bundles of the glomeruli; Tie2 was specifically expressed in endothelium of vessels both in auto- and allografts at all time points posttransplantation. At 4 weeks, the differences in mRNA expression of Ang1, Ang2, and Tie2 between the 2 groups were not significant (P > .05). Compared with autografts, the mRNA expression of Ang1 decreased significantly (P = .008 and .003 for 8 and 12 weeks, respectively), and the mRNA expressions of Ang2 and Tie2 significantly increased (P = .001/.006 and .005/.001 for 8 and 12 weeks, respectively). The changes in expression of all 3 genes showed significant correlation with the Banff score in the allografts. CONCLUSION: This study suggested that the abnormal expression and reciprocal regulation of Ang1, Ang2, and Tie2 may play important roles in the development of CAN in rat renal allografts.

Epithelial to mesenchymal transformation in tubular epithelial cells undergoing anoxia.

AIM: Epithelial-mesenchymal transformation (EMT) has been proved to be a critical event in fibrogenesis of renal allografts. This study sought to determine whether anoxia could induce EMT from tubular epithelial cells (TEC). METHODS: Rat TEC-line (NRK-52E) was cultured in Dulbelco's modified Eagle's medium (DMEM) without glucose under 100% N2 for 4 hours. After 6, 12, 24, 48, and 72 hours, the expressions of connective tissue growth factor (CTGF) mRNA and protein were measured by real-time RT-PCR and Western blot, respectively. Morphologic changes and cytoskeleton remodeling were observed in NRK-52E cells under laser confocal microscopy. Immunohistochemistry and flow cytometry were used to detect expression changes of E-cadherin, alpha-smooth muscle actin (SMA), types I and IV collagen, all of which are involved in TEC, EMT. RESULTS: After stimulation by anoxia, NRK-52E cells became round and enlarged with a remodeled cytoskeleton. The expressions of CTGF mRNA and protein were upregulated after 6 hours, reaching their peak at 48 hours. The expressions of types I and IV collagen, and alpha-SMA were all upregulated except for E-cadherin. CONCLUSIONS: Anoxia upregulated the expression of CTGF and other EMT-associated genes in NRK-52E cells.

Rapamycin and cyclosporine have different effects on expression of Ang-1 and Ang-2 and Tie2 in rat renal allograft with chronic allograft nephropathy.

OBJECTIVE: Previous studies have indicated that angiopoietin-1 (Ang1) and angiopoietin-2 (Ang2) and tyrosine kinase receptor Tie2 regulate the maintenance and integrity of blood vessels and have potential anti-inflammatory properties. This study of cardiac allografts investigated whether there is a difference between rapamycin and cyclosporine A (CsA) in the ability to affect expression of Ang1, Ang2, and Tie2 in rat renal allografts with chronic allograft nephropathy (CAN). METHODS: A male inbred F344 to Lewis rat renal CAN model was established via a modified Kamada procedure. The recipients were first treated with CsA, 10 mg/kg/d, for 10 days and then allocated randomly to three oral treatment groups: control; CsA, 6 mg/kg/d; and rapamycin, 0.8 mg/kg/d. At 4, 8, and 12 weeks posttransplantation, the rats were killed to harvest the renal allografts. The serum creatinine concentration (SCr) was measured, and the pathologic changes were assessed according to Banff 97 criteria. The expression of messengerRNA and proteins of Ang1, Ang2, and Tie2 was determined using real-time fluorescence quantitative polymerase chain reaction and immunohistochemistry. RESULTS: The elevation of SCr and the pathologic changes of CAN were observed in the control and CsA groups at 8 and 12 weeks; the differences between the 2 groups were not significant (P > .05). The levels of SCr and Banff score in the rampamycin group were lower than those in other 2 groups (P < .01). The expression of Ang1 and Ang2 was localized to epithelial cells and endothelium of vascular bundles of glomeruli, and Tie2 was specifically expressed in the endothelium of vessels in all 3 groups. At 4 weeks, the differences in mRNA expression of Ang1, Ang2, and Tie2 between 3 groups were not significant (P > .05). In a comparison of the control and CsA groups, mRNA expression of Ang1 was increased (P < .05), and mRNA expression of Ang2 and Tie2 was decreased (P < .05) in the rapamycin group at 8 and 12 weeks. The differences between the control and CsA groups were not significant at 8 or 12 weeks (P > .05). CONCLUSIONS: Our results show that compared with CsA, rapamycin modulates the expression of Ang1, Ang2, and Tie2 in rat renal allografts with CAN, which suggests that rapamycin may improve the long-term survival of renal allografts through its vasculoprotective properties.

Effects of mycophenolate mofetil on chronic allograft nephropathy by affecting RHO/ROCK signal pathways.

AIM: We sought to investigate the effects of mycophenolate mofetil (MMF) on chronic allograft nephropathy (CAN) by affecting Rho and ROCK signal pathways. METHODS: Male inbred F344 rat renal grafts orthotopically transplanted into Lewis rats were first treated with CsA (10 mg/kg(-1).d(-1) x 10 d) and then divided into 3 groups (each n = 9): (1) orally vehicle, (2) cyclosporine (CsA, 6 mg/kg(-1).d(-1)) and (3) MMF (20 mg/kg(-1).d(-1)). In addition we performed autografts of F344 (n = 10) at 4, 8, and 12 weeks, serum creatinine (SCr) was measured and pathologic changes assessed. Expression of RhoA and ROCK-1 was determined by real-time reverse transcriptase polymerase chain reaction. Expressions of alpha-smooth muscle actin (alpha-SMA) and connective tissue growth factor (CTGF) were observed by immunohistochemistry. RESULTS: SCr and Banff score began to increase at 4 weeks in all 3 allografted groups with obvious deterioration in both the vehicle and CsA groups at 8 and 12 weeks. The differences between vehicle/CsA and autografts were significant (P = .000). SCr and Banff score among the MMF group increased mildly and moderately at 8 and 12 weeks, respectively, but were significantly lower than those in the vehicle/CsA cohort (P < .05). Expressions of RhoA and ROCK-1 mRNAs and proteins were observed in mesangial and tubular cells, increasing gradually along with the progression of chronic allograft nephropathy (CAN). There was a negative correlation between RhoA/ROCK-1 mRNA and Banff score (r = -.637, p = .000; r = -.676, P = .000) or SCr (r = -.705, P = .000; r = -.756, P = .000). MMF downregulated gene and protein expressions of RhoA and ROCK-1. CsA had little effect on these expressions. Expressions of alpha-SMA and CTGF were observed in renal epithelial and tubular cells. CONCLUSION: Herein we have demonstrated abnormal expression of RhoA and ROCK-1 signal pathways, which may play roles in CAN. MMF may attenuate CAN by downregulating the expression of RhoA/ROCK-1, alpha-SMA, and CTGF.