[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.

[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.

[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.

[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.

[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.

[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.

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.

Metallothionein 2A genetic polymorphism and its correlation to coronary heart disease.

OBJECTIVE: To investigate the gene polymorphism of metallothionein 2A (MT2A)-838G/C and its correlation to coronary heart disease in the Han population of Jiangsu, China. PATIENTS AND METHODS: The MT2A-838G/C was examined in 287 patients with coronary heart disease (CHD group) and 226 healthy controls (control group) by using mono-labeled fluorescent probes. Meanwhile, relevant variables of all subjects were measured, including blood lipid and glucose profiles and body mass index (BMI). The extent of the coronary artery disease was evaluated based on Gensini's coronary artery scoring method. RESULTS: Three distinct genotypes were identified. The highest frequency was observed for genotype GG, followed by genotype GC and CC. There were statistically significant differences in the genotype and allele frequency distribution of the MT2A gene-838G/C polymorphism between the CHD and the control group (p < 0.05). The allele frequency of MT2A-838C in the CHD patients were higher than that in the healthy controls (31.4% vs 24.6%, p = 0.016). The CHD risk in C allele carriers (including genotype GC+CC) was 1.562 folds as high as in GG allele carriers (OR = 1.562, 95% confidence intervals (CI): 1.099-2.218, p = 0.013). According to the results of logistic regression analysis, the C allele was an independent risk factor for CHD (p < 0.05). Gensini's coronary artery disease score were higher in C allele carriers than in non-C allele carriers (p < 0.05). CONCLUSIONS: The gene polymorphism of MT2A-838G/C is correlated to CHD. The C allele might be a CHD-susceptible gene and might also have an effect on the extent of coronary artery disease.

Urinary apolipoprotein M could be used as a biomarker of acute renal injury: an ischemia-reperfusion injury model of kidney in rat.

BACKGROUND: It has been well documented that apolipoprotein M (apoM) is principally expressed in hepatocytes as well as renal tubular epithelial cells. The importance of apoM in the kidney is unknown. In the present study we examined urinary any apoM after short-term ischemia-reperfusion injury (IRI) of kidney in a rat model. METHODS: The kidneys of 11 male Sprague-Dawley rats were rendered ischemic for 45 minutes followed by different intervals of reperfusion. Serum and urine apoM concentrations were determined using a dot-blot analysis with specific rabbit anti-human apoM antibodies that cross-react with rat apoM. Serum concentrations of blood urea nitrogen (BUN) and creatinine (Cr) were determined using standard clinical automated analyses. RESULTS: BUN was significantly elevated after 45 minutes of ischemia followed by 24 hours of reperfusion; serum Cr concentrations were also significantly increased at 6 and 24 hours of reperfusion. Interestingly, similar to BUN and Cr, serum apoM concentrations were significantly increased after ischemia for 45 minutes alone and after 2 hours of reperfusion. Urinary apoM concentrations were obviously increased after 2 h as well as 6 hours of reperfusion. CONCLUSION: apoM showed characteristics of an acute-phase reactive protein; its occurrence in urine may be considered to be a biomarker of acute renal injury.

Urinary connective tissue growth factor is a biomarker in a rat model of chronic nephropathy.

AIM: This study sought to determine whether urinary connective tissue growth factor (CTGF) was a molecular marker for chronic allograft nephropathy (CAN). METHODS: F344 rat renal grafts orthotopically transplanted into Lewis rats following the procedure of Kamada were harvested at 4,8,12, or 16 weeks. Morphological changes were studied using hematoxylin eosin (HE) and Masson trichrome stains. Serum creatinine (SCr) was measured. CAN grades were evaluated according to the Banff97 schema. Expressions of CTGF in the kidney and urine were determined using real-time polymerase chain reaction (PCR) Western blots, and competitive indirect enzyme-linked immunosorbent assay (ELISA). Spearman correlation analysis was used to compare urinary CTGF expression and CAN development. RESULTS: SCr levels and Banff scores increased in a time-dependent manner. The expression of CTGF in the graft was markedly elevated compared with the control group. Urine CTGF increased by week 4, and maintained high levels up to week 16. The urinary levels correlated positively with the histological presence of CAN. Thus, urine CTGF concentrations reflected the course of CAN, especially at an early stage. CONCLUSION: CTGF plays a significant role in the pathological changes of CAN after kidney transplantation. Urinary CTGF has the potential to be a biomarker for CAN.

Detection of citrate synthase autoantibodies in rats with chronic allograft nephropathy.

UNLABELLED: Citrate synthase (CS) is the one of the key enzymes in the citric acid cycle and an important mitochondrial autoantigen. The autoimmune responses against CS have not been studied in chronic allograft nephropathy (CAN). This study investigated the role of specific CS autoantibodies in rats bearing renal allografts affected with CAN. METHODS: Fisher344 rat renal grafts were orthotopically transplanted into Lewis rats following the procedure of Kamada with our modification. Lewis-to-Lewis and Fisher344-to-Fisher344 kidney transplantations were also performed as autologous control groups (each n = 9). All the allograft recipients given cyclosporine (10 mg/kg(-1)d(-1) x 10 d) were divided into four groups (each n = 9): (1) vehicle: normal saline orally; (2) cyclosporine: 6 mg/kg(-1)d(-1); (3)FK506: 0.15 mg/kg(-1)d(-1); (4) mycophenolate mofetil (MMF): 20 mg/ kg(-1)d(-1). At 4, 8, and 12 weeks posttransplantation, the animals were sacrificed to harvest sera and renal allografts. The serum creatinine (SCr) was measured and pathological changes assessed according to Banff 97 criteria. IgM and IgG isotypes of CS antibodies were detected in all recipient sera by enzyme linked immunosorbent assays. RESULTS: Both IgM and IgG isotype CS autoantibodies were observed in the sera of all the recipients before and after transplantation, but the levels of IgM CS autoantibody were obviously higher than IgG isotype in all the blood samples. It was stable not only in autologous but also in allograft groups. In both autologous groups, the SCr and IgM and IgG isotype CS autoantibodies showed no obvious change before and after transplantation, and no typical CAN occurred. The values of IgG isotype of CS autoantibody (DeltaOD) at 4, 8 and 12 weeks were stable. At 4 weeks, the values of SCr, Banff score, and IgG isotype CS autoantibody (DeltaOD) were not significantly different (P > .05) among the allograft groups. At 8 and 12 weeks, with progression of CAN in vehicle, cyclosporine and FK506 groups' values of SCr, Banff score, and IgG (DeltaOD) also increased dramatically (P = .005) in all three groups when compared with the baseline and 4 week values, but the differences among the three groups were not significant (P > .05). At 8 and 12 weeks, the MMF group suffered mild-to-moderate CAN, but the values of SCr and Banff score were significantly lower than those in the other three groups. MMF significantly inhibited the formation of IgG (DeltaOD) when compared with the other three groups (P = .02). CONCLUSION: This study suggested that the IgG isotype of CS autoantibody contributes to CAN after kidney transplantation. The IgM isotype is physiological. MMF significantly inhibited the formation of IgG isotype CS autoantibody, which may be related to its effects to alleviate 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.

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.