Clinical application of "Double R" anastomosis technique in laparoscopic pancreaticoduodenectomy procedure.

ABSTRACT: Laparoscopic pancreaticoduodenectomy (LPD) is widely used as a treatment for periampullary tumors and pancreatic head tumors. However, postoperative pancreatic fistula (POPF), which significantly affects mortality and length of hospital stay of patients, remains one of the most common and serious complications following LPD. Though numerous technical modifications for pancreaticojejunostomy (PJ) have been proposed, POPF is still the "Achilles heel" of LPD.To reduce POPF rate and other postoperative complications following LPD by exploring the best approach to manage with the pancreatic remnant, a novel duct-to-mucosa anastomosis technique named Double Layer Running Suture (Double R) for the PJ was established. During 2018 and 2020, a totally 35 patients who underwent LPD with Double R were included, data on the total operative time, PJ duration, estimated blood loss, recovery of bowel function, postoperative complications, and length of hospital stay were collected and analyzed.The average duration of surgery was (380 ±â€Š69) minutes. The mean time for performing PJ was (34 ±â€Š5) minutes. The average estimated blood loss was (180 ±â€Š155) mL. The overall POPF rate was 8.6% (3/35), including 8.6% (3/35) for the biochemical leak, 0% (0/35) for Grade B, and 0% (0/35) for Grade C. No patient suffered from biliary fistula, post-pancreatectomy hemorrhage, and intra-abdominal infection, the 30-day mortality was 0%.Double R anastomosis is potentially a safe, reliable, and rapid anastomosis with a low rate of POPF and post-pancreatectomy hemorrhage. It provides surgeons more options when performing LPD. However, its safety and effectiveness should be verified further by a larger prospective multicenter study.

Comparison of transcatheter arterial chemoembolization with raltitrexed plus liposomal doxorubicin vs. tegafur plus pirarubicin for unresectable hepatocellular carcinoma.

BACKGROUND: There is still no general consensus on the optimal chemotherapeutic agent selection for transcatheter arterial chemoembolization (TACE) in unresectable hepatocellular carcinoma (HCC). The present study aimed to compare the efficacy and safety of TACE with raltitrexed plus liposomal doxorubicin (R + PGLD) vs. tegafur plus pirarubicin (T + P) in patients with unresectable HCC. METHODS: A total of 148 patients with unresectable HCC treated with TACE between January 2012 and December 2016 were retrospectively analyzed. Of them, 74 patients were in the R + PGLD group and 74 patients were in the T + P group (1:1). The treatment response of the tumor, overall survival (OS) time, and adverse effects were compared between the two groups. RESULTS: There were no significant differences in patient characteristics or embolization effect (lipiodol deposition) between the two groups (P>0.05). R + PGLD treatment had a better clinical efficacy than T + P treatment (OR: 64.9% vs. 45.9%, P=0.031; DC: 89.2% vs. 74.3%, P=0.032). Portal vein invasion, hepatic vein invasion, tumor size and BCLC stage were associated with OR or DC after TACE using R + PGLD treatment. Survival analysis revealed that patients who received TACE with R + PGLD had a better prognosis than those treated with T + P. Moreover, some complications in the R + PGLD group, including vomiting, myelosuppression and cardiotoxicity, were significantly lower than those in the T + P group (P<0.05). CONCLUSIONS: TACE with raltitrexed and liposomal doxorubicin could reduce the incidence of adverse reactions and significantly improve the OS of patients with unresectable HCC.

Oleic acid promotes cell invasion through an integrin-linked kinase signaling pathway in renal cell carcinoma.

The change of fatty acid composition has been regarded as an indicator of altered lipid metabolism during human tumourigenesis, but the details are still unclear. We have previously demonstrated a monounsaturated fatty acid (MUFA) named oleic acid (OA) was involved in renal cell carcinoma (RCC) cell growth, as an extracellular signaling molecule to regulate 786-O cell proliferation via the integrin-linked kinase (ILK) pathway. In this study, we further observe the effects of OA on cell invasion of RCC and the potential mechanism by which OA worked was determined. The transwell invasion assay showed OA increased cell invasion of RCC in a dose-dependent manner. Western blotting results indicated ILK, COX-2, and MMP-9 proteins were involved for their high expressions and these effects were reversed when down-regulating the expression of ILK by special siRNA. The MMPs inhibitor GM6001 could weaken the abilities of OA on RCC cells invasion. These results suggested MUFA indeed affected cell invasion of RCC, which was depended by the regulation of ILK pathway.

High-methionine diets accelerate atherosclerosis by HHcy-mediated FABP4 gene demethylation pathway via DNMT1 in ApoE(-/-) mice.

Homocysteine (Hcy) is an independent risk factor for atherosclerosis, but the underlying molecular mechanisms are not known. We investigated the effects of Hcy on fatty acid-binding protein 4 (FABP4), and tested our hypothesis that Hcy-induced atherosclerosis is mediated by increased FABP4 expression and decreased methylation. The FABP4 expression and DNA methylation was assessed in the aorta of ApoE(-/-) mice fed high-methionine diet for 20weeks. Over-expression of FABP4 enhanced accumulation of total cholesterol and cholesterol ester in foam cells. The up-regulation of DNA methyltransferase 1 (DNMT1) promoted the methylation process and decreased FABP4 expression. These data suggest that FABP4 plays a key role in Hcy-mediated disturbance of lipid metabolism and that DNMT1 may be a novel therapeutic target in Hcy-related atherosclerosis.

[Effect of oxymatrine on JAK2/STAT3 signaling in renal tissues of rats with septic shock].

OBJECTIVE: To explore the effect of oxymatrine (OMT) on JAK2/STAT3 signaling in renal tissues of rats with septic shock. METHOD: The cecal ligation and puncture (CLP) was adopted to establish the rat septic shock model. Fifty-six male SD rats were randomly divided into 7 groups: the sham operation group, the model (CLP) group, CLP + OMT high, middle, low-dose (52, 26, 13 mg x kg(-1), vena caudalis bolus) groups and the positive control (CLP + dexamethasone, 10 mg x kg(-1)) group. The pathological changes in renal tissues were examined with lightmicroscope. BUN content was determined by urine enzymatic method. Expressions of tumournecrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) mRNA in renal tissues were determined by RT-PCR. Expression of JAK2 and STAT3 in renal tissues determined by Western blot. Changes in tumournecrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) contents in renal tissue were determined by radioimmunoassay. RESULT: OMT of different doses could inhibit the JAK2 and STAT3 activation in renal tissues (P<0.05), and decrease the protein expression of JAK2, STAT3, TNF-alpha and IL-1beta mRNA (P<0.05). Besides, it could reduce TNF-alpha and IL-1beta contents in renal tissue homogenate (P<0.05), serum BUN content (P<0.05), and improve such lesions as tissue hyperemia, edema and inflammatory cell infiltration, with identical results in medium and high-dose OMT groups, and the positive control group. CONCLUSION: OMT can inhibit JAK2/STAT3 signaling activity to reduce the expression of proin-flammatory factors (TNF-alpha, IL-1beta) and treat the renal injury in rats with septic shock.

[Replantation of amputated penis in Chinese men: a meta-analysis].

OBJECTIVE: To evaluate the methods for the replantation of the amputated penis in Chinese men. METHODS: We performed a meta-analysis on the domestic literature relating replantation of the amputated penis, particularly its successful methods published from 1964 to January 2012. RESULTS: We identified 109 reports on 111 cases of replantation of the amputated penis that met the inclusion criteria, including 103 adults and 8 children. The mean age, warm ischemia time and total ischemia time were 29 +/- 11 years (range 2 - 56 years), 5.2 +/- 5.7 hours (range 0 - 38 hours) and 6.3 +/- 5.7 hours (range 1 - 38 hours). Fifty-three of the cases were treated by microsurgery and 44 by non-microsurgery. Complications occurred in 81 (73%) of the cases, including ED in 14 cases, urethral stricture in 16, urinary fistula in 8, skin necrosis in 58 and skin sensory abnormality in 31. The incidences of ED, urethral stricture and urinary fistula exhibited significant differences between the microsurgery and non-microsurgery groups of the partial amputation patients (P < 0.05). The incidence of ED was correlated negatively with the number of anastomosed dorsal nerves (r = -0.3, P = 0.05), anastomosis of dorsal veins (r = -0.2, P = 0.02) and anastomosis of arteries (r = -0.2, P = 0.03), but positively with skin sensory abnormality (r = 0.4, P < 0.01), that of urethral stricture negatively with the anastomosis of dorsal nerves (r = -0.2, P = 0.02) and arteries (r = -0.2, P = 0.016), but positively with the anastomosis of corpus cavernosum (r = 0.3, P = 0.01), that of skin necrosis negatively with the total number of anastomosed blood vessels (r = -0.2, P = 0.04), and that of complications negatively with the number of anastomosed dorsal nerves (r = -0.3, P = 0.01), dorsal veins (r = -0.2, P = 0.04), arteries (r = -0.2, P = 0.023) and micro-anastomosis (r = -0.3, P < 0.05). CONCLUSION: Early micro-anastomosis of the most possible penile dorsal veins, arteries and dorsal nerves is essential for the survival of the replanted penis and reduction of complications, and therefore can be regarded as a "standard" method for penile replantation in China.

Multiple factors related to detrusor overactivity in Chinese patients with benign prostate hyperplasia.

BACKGROUND: Detrusor overactivity (DO) is a known cause of lower urinary tract symptoms and occurs in 50% - 75% of benign prostate hyperplasia (BPH) patients. We sought to investigate the clinical and urodynamic factors that are associated with the presence of DO in Chinese BPH patients. METHODS: Two hundred and eighty-seven consecutive patients with clinical BPH were retrospectively evaluated in this study. Each patient underwent urodynamic evaluation and completed the International Prostate Symptom Score (IPSS) and Quality of Life (QoL) questionnaire. Patients with neurological symptoms or other diseases likely to affect detrusor functions were strictly excluded. The 184 BPH patients included in the study were divided into groups according to the presence of DO as shown in urodynamic tests. Univariate analysis of factors associated with the presence of DO were performed using Student's t-test and the Mann-Whitney test; multivariate analysis used stepwise Logistic regressions. The relationship between degree of bladder outlet obstruction (BOO) and DO was also investigated using a linear-by-linear association test. RESULTS: Of 184 BPH patients, DO was present in 76 (41.3%). On univariate analysis, patients with DO were older (P = 0.000), and showed smaller maximal bladder capacity (MBC, P = 0.000) and voided volume (P = 0.000), higher maximal detrusor pressure (P = 0.000) and projected isovolumetric pressure (PIP) (P = 0.005), higher Abrams-Griffiths number (P = 0.000) and degree of bladder outlet obstruction (P = 0.000), higher IPSS (P = 0.000) and irritative IPSS subscores (P = 0.000). Stepwise Logistic regression analysis showed that PIP (OR = 1.012, 95% CI 1.002 - 1.023, P = 0.019), age (OR = 1.030, 95%CI 1.005 - 1.067, P = 0.059), and MBC (OR = 0.993, 95%CI 0.990 - 0.996, P = 0.000) were independent risk factors for DO in BPH patients. Linear-by-linear association tests indicated a positive linear association between DO and severity of BOO, with incidence of DO increasing with BOO grade (P = 0.000). CONCLUSIONS: In Chinese BPH patients, PIP, MBC, and age were independent factors affecting the presence of DO. DO incidence continuously increases with the degree of BOO.

[Aggressiveness and extent of prostatic inflammation relates with serum PSA levels in type IV prostatitis].

OBJECTIVE: To investigate the relationship of the histopathologic grade and extent of prostatic inflammation with the level of serum PSA in patients with type IV prostatitis. METHODS: We performed transrectal ultrasound-guided prostate biopsy for 120 patients suspected of prostate cancer and included in this study only those with benign prostate hyperplasia (BPH) and prostatitis (n = 46), excluding the cases with prostate cancer and those with BPH but no prostatitis. We evaluated the relationship between prostatic inflammation and serum PSA levels based on the three-grade pathohistologic criteria for the extent, location and aggressiveness of prostatic inflammation. The serum tPSA levels, fPSA levels, % fPSA, and PSAD were compared among different groups. RESULTS: As for the extent of inflammation, 35 of the 46 included cases were grade I (tPSA: [8.46 +/- 4.09] microg/L; fPSA: [1.75 +/- 0.93] microg/L; PSAD: 0.15 +/- 0.11), 7 were grade II (tPSA: [15.26 +/- 5.26] microg/L; fPSA: [2.54 +/- 0.72] microg/L; PSAD: 0.26 +/- 0.07) and 4 were grade III (tPSA: [21.05 +/- 7.58] microg/L; fPSA: [3. 19 +/- 1.13] microg/L; PSAD: 0.42 +/- 0.19), with statistically significant differences among the three groups in the levels of tPSA (P = 0.001), fPSA (P = 0.008) and PSAD (P < 0.001). Regarding the location of inflammation, 19 cases were grade I, 17 were grade II and 10 were grade II, with no significant differences in tPSA, fPSA and %fPSA among the three grades (P > 0.05). As for the aggressiveness of inflammation, 32 cases were grade I (tPSA: [8.37 +/- 4.07] microg/L; fPSA: [1.76 +/- 0.93] microg/L; PSAD: 0.14 +/- 0.11), 10 were grade II (tPSA: [13.30 +/- 5.69] microg/L; fPSA: [3.27 +/- 2.21] microg/L ; PSAD: 0.25 +/- 0.06) and 4 were grade III (tPSA: [21.05 +/- 7.58] microg/L; fPSA: [3.19 +/- 1.13] microg/L; PSAD: 0.42 +/- 0.19), with statistically significant differences among the three grades in the levels of tPSA (P = 0.002), fPSA (P = 0.024) and PSAD (P < 0.001). The extent of inflammation was positively correlated with the levels of tPSA (r = 0.6, P < 0.001), fPSA (r = 0.5, P = 0.001) and PSAD (r = 0.6, P < 0.001), and so was the aggressiveness of inflammation (tPSA: r = 0.5, P < 0.001; fPSA: r = 0.4, P = 0.008; PSAD: r = 0.7, P < 0.001), but a negative correlation was found between the aggressiveness of inflammation and %fPSA (r = -0.4, P = 0.013). CONCLUSION: The aggressiveness and extent of prostatic inflammation in asymptomatic prostatitis patients are significantly correlated with the level of serum PSA, which may help pathologists to avoid unnecessary repeated biopsies for patients with high-grade prostatitis.

Risk factors for interstitial cystitis/painful bladder syndrome in patients with lower urinary tract symptoms: a Chinese multi-center study.

BACKGROUND: Despite 100 years of research, the continued absence of well-established risk factors impedes the diagnosis and treatment of interstitial cystitis/painful bladder syndrome (IC/PBS). We aimed to identify risk factors in patients with lower urinary tract symptoms (LUTS) without urinary tract infection or benign prostate hyperplasia in China. METHODS: A total of 397 outpatients with LUTS presenting for care to urology clinics in several hospitals throughout China were surveyed using a standardized questionnaire and validated outcome measures. The definitions for painful bladder syndrome based on the O'Leary-Sant interstitial cystitis symptom and problem indices were used. The prevalence of possible risk factors was analyzed using the Fisher's exact test and Pearson chi-square test, and multivariate predictive models were developed using binary Logistic regression methods. RESULTS: Of those multi-centre patients surveyed, including 174 women and 223 men, 41% (162/397) met criteria for painful bladder syndrome. There was a significant difference between women and men (55% (95/174) vs. 30% (67/223), P < 0.001). Women with IC/PBS were more likely than those without IC/PBS to report a history of gynecological infections (odds ratio (OR): 2.85; 95% confidence interval (CI): 1.32 - 6.16, P = 0.007), intake of stimulatory foods (OR: 3.52; 95%CI: 1.50 - 8.30; P = 0.004), irritable bowel (OR: 3.46; 95%CI: 1.22 - 9.80; P = 0.014) and/or anorectal disease (OR: 2.68; 95%CI: 1.12 - 6.40, P = 0.023). After adjusting for confounding factors, bladder pain was significantly associated with stimulatory foods (OR: 3.85; 95%CI: 1.58 - 9.36, P = 0.003) and anorectal disease (OR: 2.76; 95%CI: 1.09 - 7.04, P = 0.03) in women. Caffeine beverage intake (OR: 3.54; 95%CI: 1.54 - 8.12, P = 0.003) was identified the only modifiable association noted in multivariate analysis of men. CONCLUSIONS: We found that stimulatory foods, anorectal disease and caffeine beverages are potential risk factors for IC/PBS. Further studies are necessary to determine their role in the pathogenesis of this disorder.

[Analysis of relation between types of bony metastasis and PSA and tumor differentiation of prostate cancer].

OBJECTIVE: To analyze the relationship between radiological characteristics of bony metastasis and PSA, Gleason score (GS) for prostate cancer patients. METHODS: The investigators collected the clinical data of 44 patients with prostate cancer bony metastasis and divided them into 3 groups based upon the radiological characteristics of bony metastasis. The 3 groups were osteoblast group, osteoclast group and mixed group. RESULTS: The mean age of patients was 70 years old. There were 33 cases (75.00%), 9 cases (20.45%) and 2 cases (4.55%) in osteoblast group, osteoclast group and mixed group respectively. The mean total serum PSA values were 590.3+/-177.0 microg/L, 249.0+/-66.6 microg/L and 156.5+/-42.1 microg/L in osteoblast group, osteoclast group and mixed group respectively. No statistic significance was found for the difference of total PSA in 3 groups. Mean GS in osteoblast group was 7.4 (6-10). Twenty-two cases (66.7%) were of a medium-high differentiation (GS7). Mean GS in osteoclast group was 8.0 (6-10). Among which, 3 cases (33.3%) were of a medium-high differentiation while 6 cases (66.7%) of a low differentiation. Statistic significance was found for the difference of GS in 2 groups. Two cases in mixed group were assigned 8 of GS. CONCLUSION: There is no confirmed correlation between types of bony metastasis and total serum PSA for prostate cancer patients. But the tumor differentiation reflected by Gleason score may affect the type of bony metastasis.

[Clinical comparison of flexible endoscopic realignment with traditional open realignment].

OBJECTIVE: To evaluate the clinical value of flexible endoscopic realignment for posterior urethral disruption as compared with traditional open realignment. METHODS: A total of 58 patients suffering posterior urethral disruption in Beijing Jishuitan Hospital from January 2003 to May 2009 were examined. Twenty-three patients (Group A) were performed with flexible endoscopic realignment and 35 patients (Group B) were performed with traditional open realignment. The operation duration and post-operative complications were compared between two groups. RESULTS: All operations were successfully performed. The operative duration was significantly shorter in Group A than that in Group B [(29 +/- 10) min vs (58 +/- 11) min, P = 0.000]. Also, the patients in Group A had a significantly decreased rate of stricture and false urethral passages formation (17.4% vs 42.9%, 0 vs 20%, both P < 0.05). There were no significant difference in urinary infection, repeat procedures, incontinence and impotence between two groups (P > 0.05). CONCLUSION: As compared with traditional open realignment, flexible endoscopic realignment has the advantages of short operation duration, lesser trauma and fewer complications. Therefore it is an excellent method for the treatment of urethral injury.

[Impact of disease duration of lumbar disk herniation on bladder dysfunction].

OBJECTIVE: To analyze the association between the disease duration and bladder dysfunction in lumbar disk herniation patients. METHODS: The urodynamic parameters of 85 lumbar disk herniation patients with different disease duration, 26 with the duration < 1 year (Group A), 31 with the duration of 1 - 5 years (Group B), and 28 of the duration > 5 years (Group C), were retrospectively analyzed. RESULTS: The average post-voiding residual volume of Group A was 50 ml, significantly lower than those of Groups B and C (80 ml and 120 ml respectively, both P < 0.05) with significant difference between Groups B and C too (P < 0.05). The average maximum cystometric capacity of Group A was (375 +/- 147) ml, significantly higher than those of Groups B and C [(481 +/- 184) ml and (562 +/- 173) ml respectively, both P < 0.05] and there was significant difference between Groups B and C too (P < 0.05). The average volume at the first desire to void of Group A was (220 +/- 88) ml, significantly lower than those of Groups B and C [(247 +/- 132) ml and (356 +/- 199) ml respectively, both P < 0.05], however, there was no significant difference between Groups A and B (P > 0.05). There were no significant differences in the maximum flow rate, maximum detrusor pressure, and bladder compliance among the three groups (all P > 0.05). CONCLUSION: Prolonged disease duration of lumbar disk herniation worsens the bladder function.

[Effect of oxymatrine on NF-kappaB and other cell factors in rats lung tissue with septic shock].

OBJECTIVE: To explore the effects of oxymatrine (OMT) on NF-kappaB and other cell factors in rat lung tissue with septic shock. METHOD: Fifty-six male SD rats were randomly divided into 7 groups: sham operation group, OMT control group, model (CLP) group, CLP + OMT high, middle, low-dose group, positive control group. Changes in NF-kappaB (p65) and IkB-alpha activity in the pulmonary tissue were determined by immunohistochemical method, tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) levels in pulmonary tissue were determined by radioimmunoassay. RESULT: OMT could decrease significantly the NF-kappaB (p65) and IkB-alpha activity in the pulmonary tissue (P < 0.05), TNF-alpha and IL-6 levels in pulmonary tissue homogenate decreased markedly (P < 0.05). OMT could elevate the content of PaO2, SaO2, decrease the content of PaCO2, HCO3- and decrease the ratio between wet weight of the lung and dry weight of the lung and the PWI. CONCLUSION: OMT can inhibit NF-kappaB-inducing kinase (NIK), NF-kappaB activity and reduce the expression of proinflammatory factor (TNF-alpha, IL-6) and antagonize the lung injury in a rat model of septic shock.

[Clinical value of ultrasound guided transperineal prostate biopsy in detecting prostate cancer].

OBJECTIVE: To report our experience of ultrasound guided transperineal 6-core prostate biopsy (UG6CPB) in the diagnosis of prostate cancer (PCa). METHODS: In a prospective study, we performed UG6CPB in 104 suspected PCa patients with tPSA more than 4 microg/L and analysed the positive rate and complications of the diagnostic approach. RESULTS: PCa was detected in 24 of the 104 patients (23%), with low grade Gleason 2 to 4 in 3 cases (12.5%), intermediate grade Gleason 5 to 7 in 15 (62.5%) and high grade Gleason 8 to 10 in the remaining 6 (25%). Complications included temporary hematuria in 5 patients (4.8%), mild postbiopsy perineal discomfort in 5 (4.8%) and fever in 4 (3.8%). TPSA > or =10 microg/L, fPSA > or = 2 microg/L, fPSA/tPSA < 0.16, PSAD > or = 0.2 and prostate volume < 40 ml were the significant influencing factors of biopsy positive rate (P < 0.05). CONCLUSION: UG6CPB is an exact and a safe way of detecting PCa.

Ghrelin blunted vascular calcification in vivo and in vitro in rats.

Ghrelin is a new peptide with regulatory actions in growth hormone secretion in the anterior pituitary gland and in energy metabolism. Currently, ghrelin has potently protective effects in cardiovascular diseases. We used an in vivo model of rat vascular calcification induced by vitamin D3 and nicotine and one of cultured rat vascular smooth muscular cells (VSMCs) calcification induced by beta-glycerophosphate to study the possible mechanism in the regulatory action of ghrelin in vascular calcification. Calcification increased total Ca2+ content and 45Ca2+ deposition in aortas and VSMCs and alkaline phosphatase (ALP) activation in plasma, aortas and VSMCs. However, calcified aortas and VSMCs showed a significant decrease in osteopontin (OPN) mRNA expression and a marked reduction of ghrelin levels in plasma and its mRNA expression in aortas. The aortic calcification was significantly attenuated by subcutaneous administration of ghrelin 30 and 300 nmol kg(-1) day(-1) for 4 weeks, and the latter dosage was more potent than the former. Ghrelin treatment at the two dosages reduced the total aorta Ca2+ content by 24.4% and 28.1%, aortic 45Ca2+ deposition by 18.4% and 24.9%, plasma ALP activity by 36.6% and 76.7%, and aortic ALP activity by 10.3% and 47.6% (all P < 0.01 or 0.05), respectively. Ghrelin at 10(-8)-10(-6) mol/L attenuated the calcification in cultured VSMCs, with decreased total Ca2+ content, 45Ca2+ deposition and ALP activity and increased OPN mRNA expression, in a concentration-dependent manner. In addition, endothelin levels in plasma and aortas and its mRNA expression in aortas significantly increased with calcification, but ghrelin treatment significantly decreased endothelin levels and mRNA expression, with the high dosage being more potent than the lower dosage. These results indicate that local ghrelin in vascular was down-regulated during vascular calcification, whereas administration of ghrelin effectively attenuated vascular and VSMCs calcification.

Ghrelin protects myocardium from isoproterenol-induced injury in rats.

AIM: To investigate the cardiac protective effects of ghrelin in rat with myocardial injury induced by isoproterenol (ISO). METHODS: Rats were subcutaneously injected ISO 40 mg/kg/d with or without ghrelin 1 or 10 nmol/kg/d for 2 d. Hemodynamic parameters including mean arterial blood pressure and left ventricular pressure were measured at 12 h after the last injection with ISO and/or ghrelin. Plasma lactate dehydrogenase (LDH) activity, plasma and myocardial contents of malondialdehyde (MDA), and conjugated diene were measured. Plasma ghrelin and endothelin-1 levels were assayed using radioimmunoassay methods. Endothelin-1 and ghrelin mRNA were determined using RT-PCR. RESULTS: About 45 % (5/11) of rats after treatment with ISO alone died during experimental periods. However, no rats died after administration with ghrelin 10 nmol/kg/d (0/11, P<0.05). Ghrelin also obviously ameliorated the hemodynamic disturbance in rats induced by ISO. The plasma LDH activity, contents of myocardial and plasma MDA, and conjugated diene level in plasma in ISO+G10 nmol/kg/d group were decreased by 28 %, 34 %, 73 %, and 38 % compared with those of ISO group (all P<0.01) respectively. ISO-induced endothelin-1 mRNA over-expression was inhibited and endothelin-1 level in plasma were inhibited by ghrelin 1 and 10 nmol/kg/d. The ghrelin levels in plasma and ghrelin mRNA in myocardium were increased in the rats after injection of ISO. The plasma ghrelin level was further increased after ghrelin administration. CONCLUSION: Ghrelin has a protective effect against ISO-induced myocardial injury.

Alterations of adrenomedullin and its receptor system components in calcified vascular smooth muscle cells.

Vascular calcification is a common finding in many cardiovascular diseases. Paracrine/autocrine changes in calcified vessels, and the secreted factors participate in and play an important role in the progress of calcification. Adrenomedullin (ADM) is a potent vasodilator peptide secreted by vascular smooth muscle cells (VSMCs) and vascular endothelial cells. Recently, receptor activity-modifying proteins (RAMPs) have been shown to transport calcitonin receptor-like receptor (CRLR) to the cell surface to present either as CGRP receptor or ADM receptor. In this work, we explored the production of ADM, alterations and significance of ADM mRNA and its receptor system components--CRLR and RAMPs mRNA in calcified VSMCs. Our results showed that calcium content, 45Ca2+ uptake and alkaline phosphatases (ALPs) activity in calcified VSMCs were increased, respectively, compared with control VSMCs. Content of ADM in medium was increased by 99% (p < 0.01). Furthermore, it was found that the levels of ADM, CRLR, RAMP2 and RAMP3 mRNA in calcified cells were elevated, respectively, compared with that of control. The elevated levels of CRLR, RAMP2 and RAMP3 mRNA were significant correlation with ADM mRNA (r = 0.83, 0.92 and 0.93, respectively, all p's < 0.01) in calcified VSMCs. The results show that calcified VSMCs generate an increased amount of ADM, up-regulate gene expressions of ADM and its receptor system components--CRLR, RAMP2 and RAMP3, suggesting an important role of ADM and its receptor system in the regulation of vascular calcification.

[Alterations of adrenomedullin and its receptor system in calcified myocardium and aorta of rats].

OBJECTIVE: To explore the production of ADM, changes and significance of adrenomedullin (ADM) mRNA and ADM receptor system-calcitonin receptor-like receptor (CRLR) and receptor activity modifying proteins (RAMPs) mRNA in calcified myocardium and aorta of rats. METHODS: Contents of ADM in plasma, myocardium and vessel were measured by radioimmunoassay (RIA). The amount of ADM, CRLR and RAMPs mRNA was determined by semi-quantitative RT-PCR. RESULTS: The contents of calcium in calcified myocardium and aorta were increased by 342% and 606% (all P<0.01), respectively, and alkaline phosphatase activities in calcified myocardium and vessel were increased by 66.5% and 82.7% (all P<0.01), respectively, compared with the control. Contents of ADM in plasma, myocardium and aorta were increased by 58 % (P<0.01), 14.3%(P<0.01) and 27.8%(P<0.05). Furthermore, it was found that the levels of ADM, CRLR and RAMP2 mRNA in calcified myocardium were elevated by 90.6% 157.5% and 119.6%(all P<0.01), respectively, but RAMP3 mRNA was decreased by 14.1 %(P<0.01). The levels of ADM, CRLR, RAMP2 and RAMP3 mRNA in calcified aorta were elevated by 37.7%(P<0.01), 41.4%(P<0.01),60.1%(P<0.05) and 13%(P<0.01) respectively, compared with the control. The elevated levels of CRLR, RAMP2 or RAMP3 mRNA were positively correlated with up regulations of ADM mRNA in calcified myocardium and aorta. CONCLUSION: Calcified myocardium and aorta may generate an increased amount of ADM, different alterations of gene expressions of ADM and its receptor system. These RESULTS suggest that ADM and its receptor system might participate in the regulation of calcification in heart and vessel.