Hong-Hwa Chen

Additional benefit of combined therapy with melatonin and apoptotic adipose-derived mesenchymal stem cell against sepsis-induced kidney injury

This study tested whether combined therapy with melatonin and apoptotic adipose‐derived mesenchymal stem cells (A‐ADMSCs) offered additional benefit in ameliorating sepsis‐induced acute kidney injury. Adult male Sprague–Dawley rats (n = 65) were randomized equally into five groups: Sham controls (SC), sepsis induced by cecal‐ligation and puncture (CLP), CLP‐melatonin, CLP‐A‐ADMSC, and CLP‐melatonin‐A‐ADMSC. Circulating TNF‐α level at post‐CLP 6 hr was highest in CLP and lowest in SC groups, higher in CLP‐melatonin than in CLP‐A‐ADMSC and CLP‐melatonin‐A‐ADMSC groups (all P < 0.001). Immune reactivity as reflected in the number of splenic helper‐, cytoxic‐, and regulatory‐T cells at post‐CLP 72 hr exhibited the same pattern as that of circulating TNF‐α among all groups (P < 0.001). The histological scoring of kidney injury and the number of F4/80+ and CD14+ cells in kidney were highest in CLP and lowest in SC groups, higher in CLP‐melatonin than in CLP‐A‐ADMSC and CLP‐melatonin‐A‐ADMSC groups, and higher in CLP‐A‐ADMSC than in CLP‐melatonin‐A‐ADMSC groups (all P < 0.001). Changes in protein expressions of inflammatory (RANTES, TNF‐1α, NF‐κB, MMP‐9, MIP‐1, IL‐1β), apoptotic (cleaved caspase 3 and PARP, mitochondrial Bax), fibrotic (Smad3, TGF‐β) markers, reactive‐oxygen‐species (NOX‐1, NOX‐2), and oxidative stress displayed a pattern identical to that of kidney injury score among the five groups (all P < 0.001). Expressions of antioxidants (GR+, GPx+, HO‐1, NQO‐1+) were lowest in SC group and highest in CLP‐melatonin‐A‐ADMSC group, lower in CLP than in CLP‐melatonin and CLP‐A‐ADMSC groups, and lower in CLP‐melatonin‐ than in CLP‐A‐ADMSC‐tretaed animals (all P < 0.001). In conclusion, combined treatment with melatonin and A‐ADMSC was superior to A‐ADMSC alone in protecting the kidneys from sepsis‐induced injury.

Protective effect of melatonin-supported adipose-derived mesenchymal stem cells against small bowel ischemia-reperfusion injury in rat

We tested the hypothesis that combined melatonin and autologous adipose‐derived mesenchymal stem cells (ADMSC) was superior to either alone against small bowel ischemia‐reperfusion (SBIR) injury induced by superior mesenteric artery clamping for 30 min followed by reperfusion for 72 hr. Male adult Sprague Dawley rats (n = 50) were equally categorized into sham‐operated controls SC, SBIR, SBIR‐ADMSC (1.0 × 106 intravenous and 1.0 × 106 intrajejunal injection), SBIR‐melatonin (intraperitoneal 20 mg/kg at 30 min after SI ischemia and 50 mg/kg at 6 and 18 hr after SI reperfusion), and SBIR‐ADMSC‐melatonin groups. The results demonstrated that the circulating levels of TNF‐α, MPO, LyG6+ cells, CD68+ cells, WBC count, and gut permeability were highest in SBIR and lowest in SC, significantly higher in SBIR‐ADMSC group and further increased in SBIR‐melatonin group than in the combined therapy group (all P < 0.001). The ischemic mucosal damage score, the protein expressions of inflammation (TNF‐α, NF‐κB, MMP‐9, MPO, and iNOS), oxidative stress (NOX‐1, NOX‐2, and oxidized protein), apoptosis (APAF‐1, mitochondrial Bax, cleaved caspase‐3 and PARP), mitochondrial damage (cytosolic cytochrome C) and DNA damage (γ‐H2AX) markers, as well as cellular expressions of proliferation (PCNA), apoptosis (caspase‐3, TUNEL assay), and DNA damage (γ‐H2AX) showed an identical pattern, whereas mitochondrial cytochrome C exhibited an opposite pattern compared to that of inflammation among all groups (all P < 0.001). Besides, antioxidant expressions at protein (NQO‐1, GR, and GPx) and cellular (HO‐1) levels progressively increased from SC to the combined treatment group (all P < 0.001). In conclusion, combined melatonin‐ADMSC treatment offered additive beneficial effect against SBIR injury.

Melatonin treatment further improves adipose-derived mesenchymal stem cell therapy for acute interstitial cystitis in rat.

This study tests the hypothesis that combined melatonin and adipose‐derived mesenchymal stem cell (ADMSC, 1.2 × 106 given intravenously) treatment offer superior protection against cyclophosphamide (CYP 150 mg/kg)‐induced acute interstitial cystitis (AIC) in rats. Male adult Sprague‐Dawley rats were treated as follows: sham controls, AIC alone, AIC + melatonin, AIC + ADMSC, and AIC + melatonin +ADMSC. When melatonin was used, it was given as follows: 20 mg/kg at 30 min after CYP and 50 mg/kg at 6 and 18 hr after CYP. Twenty‐four‐hour urine volume, urine albumin level, and severity of hematuria were highest in AIC rats and lowest in the controls; likewise urine volume was higher in AIC + melatonin rats than in AIC + ADMSC and AIC + melatonin + ADMSC treated rats; in all cases, P < 0.001. The numbers of CD14+, CD74+, CD68+, MIP+, Cox‐2+, substance P+, cells and protein expression of IL‐6, IL‐12, RANTES, TNF‐α, NF‐κB, MMP‐9, iNOS (i.e. inflammatory biomarkers), glycosaminoglycan level, expression of oxidized protein, and protein expression of reactive oxygen species (NOX‐1, NOX‐2, NOX‐4) in the bladder tissue exhibited an identical pattern compared with that of hematuria among the five groups (all P < 0.0001). The integrity of epithelial layer and area of collagen deposition displayed an opposite pattern compared to that of hematuria among all groups (P < 0.0001). The cellular expressions of antioxidants (GR, GPx, HO‐1, NQO 1) showed a significant progressive increase form controls to AIC + melatonin + ADMSC (all P < 0.0001). Combined regimen of melatonin and ADMSC was superior to either alone in protecting against CYP‐induced AIC.

Melatonin pretreatment enhances the therapeutic effects of exogenous mitochondria against hepatic ischemia-reperfusion injury in rats through suppression of mitochondrial permeability transition.

We tested the hypothesis that melatonin (Mel) enhances exogenous mitochondria (Mito) treatment against rodent hepatic ischemia–reperfusion (IR) injury. In vitro study utilized three groups of hepatocytes (i.e. nontreatment, menadione, and menadione–melatonin treatment, 4.0 × 105 each), while in vivo study used adult male Sprague Dawley rats (n = 40) equally divided into sham‐control (SC), IR (60‐min left‐lobe ischemia + 72‐hr reperfusion), IR‐Mel (melatonin at 30 min/6/8 hr after reperfusion), IR‐Mito (mitochondria 15,000 μg/rat 30 min after reperfusion), and IR‐Mel‐Mito. Following menadione treatment in vitro, oxidative stress (NOX‐1/NOX‐2/oxidized protein), apoptotic (cleaved caspase‐3/PARP), DNA damage (γ‐H2AX/CD90/XRCC1), mitochondria damage (cytosolic cytochrome c) biomarkers, and mitochondrial permeability transition were found to be lower, whereas mitochondrial cytochrome c were found to be higher in hepatocytes with melatonin treatment compared to those without (all P < 0.001). In vivo study demonstrated highest liver injury score and serum AST in IR group, but lowest in SC group and higher in IR‐Mito group than that in groups IR‐Mel and IR‐Mel‐Mito, and higher in IR‐Mel group than that in IR‐Mel‐Mito group after 72‐hr reperfusion (all P < 0.003). Protein expressions of inflammatory (TNF‐α/NF‐κB/IL‐1β/MMP‐9), oxidative stress (NOX‐1/NOX‐2/oxidized protein), apoptotic (caspase‐3/PARP/Bax), and mitochondria damage (cytosolic cytochrome c) biomarkers displayed an identical pattern, whereas mitochondria integrity marker (mitochondrial cytochrome c) showed an opposite pattern compared to that of liver injury score (all P < 0.001) among five groups. Microscopically, expressions of apoptotic nuclei, inflammatory (MPO+/CD68+/CD14+ cells), and DNA damage (γ‐H2AX+ cells) biomarkers exhibited an identical pattern compared to that of liver injury score (all P < 0.001) among five groups. Melatonin‐supported mitochondria treatment offered an additional benefit of alleviating hepatic IR injury.

Apoptotic adipose-derived mesenchymal stem cell therapy protects against lung and kidney injury in sepsis syndrome caused by cecal ligation puncture in rats

IntroductionWe tested the hypothesis that apoptotic adipose-derived mesenchymal stem cells (A-ADMSC) are superior to healthy (H)-ADMSC in attenuating cecal ligation puncture (CLP)-induced sepsis-mediated lung and kidney injuries.MethodsAdult male rats divided into group 1 (sham controls), group 2 (CLP), group 3 [CLP + H-ADMSC administered at 0.5, 6, and 18 hours after CLP], and group 4 [CLP + A-ADMSC administered as in group 3] were sacrificed 72 hours after CLP with blood, lung, and kidney collected for studies.ResultsWhite blood cell (WBC) count, circulating TNF-α and creatinine levels were higher in groups 2 and 3 than in groups 1 and 4 (all P < 0.001). Kidney and lung damage scores were highest in group 2, lowest in group 1, significantly higher in group 3 than in group 4 (all P < 0.0001). Protein expressions of inflammatory (ICAM-1, MMP-9, TNF-α, NF-κB), oxidative, and apoptotic (Bax, caspase-3, PARP) biomarkers were higher in groups 2 and 3 than groups 1 and 4, whereas anti-apoptotic (Bcl-2) and mitochondrial integrity (cytochrome-C) biomarkers were lower in groups 2 and 3 than in groups 1 and 4 (all P < 0.001). Expressions of anti-oxidant biomarkers at protein (GR, GPx, NQO-1, HO-1) and cellular (GR, GPx) levels were highest in group 4 (all P < 0.001). The number of inflammatory cells (CD3+) in lungs and levels of DNA damage marker (γ-H2AX) in kidneys were higher in groups 2 and 3 than in groups 1 and 4 (all P < 0.001).ConclusionsA-ADMSC therapy was superior to H-ADMSC therapy in protecting major organs from damage in rats with CLP-induced sepsis syndrome.

Lipocalin 2 Level Predicts Prognostic Outcome in Patients with Colorectal Cancer Undergoing Surgical Intervention

This study examined the predictive role of circulating lipocalin 2 in long-term prognostic outcome of Colorectal Cancer (CRC) patients with surgical intervention. 204 CRC patients with surgical intervention between April 2009 and May 2011 were prospectively enrolled. Circulating lipocalin 2 levels in CRC patients before surgery and healthy-control subjects were measured using ELISA. Patients were categorized into different CRC clinical stage (I to IV) based on pathological findings. Mean follow-up time was 21.2 ± 3.5 months. Patients were categorized into group 1 (n=67), lipocalin 2 levels <116.7 ng/ ml; and group 2 (n=137), lipocalin 2 levels ≥ 116.7 ng/ml by ROC curve analysis. CRC patients had higher circulating levels of lipocalin 2 than healthy subjects (P<0.0001). Occurrence of new metastases after surgery, primary (any cause of death or new metastasis) and secondary (CRC-related death and new metastasis) combined endpoint were higher in group 1 than group 2 (all P<0.03). Lower lipocalin 2 levels and CRC stage IV was independently predictive of primary and secondary combined endpoint (all P<0.001) during long-term follow-up. In conclusion, circulating lipocalin 2 level may be a useful biomarker for risk stratification of CRC patients during long-term follow-up.

Surgical Management of Complete Rectal Prolapse

Purpose. To review our experience in the surgical management of complete rectal prolapse. Methods. Medical records of 40 patients (16 males, 24 females; mean age 59.6 20.7 years; range 17-86 years) who received surgical treatment of complete rectal prolapse from January 1997 to December 2006 were reviewed retrospectively. Results. Patients in the abdominal group (n=28) underwent recto-went laparoscopic with or without rectopexy, of which eight patients underwent laparoscopic surgery. No postoperative mortality occurred. Major perioperative complications included one anastomotic leakage in both the abdominal group and the perineal group (n=12). Recurrence rates were 2 of 28 (7.2%) in the abdominal group and 4 of 12 (33.3%) in the perineal group; p=0.034. Only 2 of the latter 4 patients received surgical treatment for the recurrent rectal prolapse (one abdominal procedure and one repeated perineal procedure). The latter patient experienced internal bleeding and was converted to laparotomy. Conclusions. An abdominal procedure can have a significantly lower recurrence rate than the perineal procedure, and so is recommended for younger and healthier patients. The perineal procedure, which has a higher recurrence rate but lower surgical risk, is more suitable for elderly patients or those with higher operative risk.

Impact of rosuvastatin treatment on reduction of thrombus burden in rat acute inferior vena cava stenosis

BackgroundThis study tested the hypothesis that rosuvastatin reduces thrombus burden through inhibiting inflammation and suppressing reactive oxygen species (ROS) generation in an inferior vena cava stenosis (IVCST)-induced deep vein thrombosis (DVT) rat model.Methods12-week-old male Sprague-Dawley rats (n = 24) were equally divided into sham control (group 1: laparotomy only), IVCST (group 2: IVC stenosis), and IVCST + rosuvastatin (20 mg/kg/day, orally after induction of IVC stenosis) (group 3). IVC diameter was measured by days 0 and 14 and the right hindlimb thickness was measured by day 0, 7, and 14 prior to scarifying the animals.ResultsThe results showed significantly increased IVC diameter and hindlimb thickness in group 2 than in groups 1 and 3, and significantly increased in group 3 than in group 1 by day 14 after the procedure (all p < 0.001). Additionally, WBC count and prevalence of helper T cells, cytotoxic T cells, regulatory T cells, and early and late apoptotic mononuclear cells (MNCs) in circulation were significantly higher in group 2 than in group 1, and were significantly suppressed in group 3 after treatment (all p < 0.001). Furthermore, inflammation at cellular (CD68+ cells) and protein (MMP-9, TNF-α) levels, oxidative stress (oxidized protein) and reactive oxygen species (NOX-1, NOX-2) in IVC also showed similar changes as those of immune cells in circulation among the three groups (all p < 0.01).ConclusionRosuvastatin treatment significantly reduced IVC thrombus burden through inhibiting inflammatory response and oxidative stress in a rodent model of DVT.

Treatment Response in Patients with Primary Colonic Lymphomas: Experience in a Single Institute

Purpose. To review the clinical characteristics, treatments and prognosis of the rare malignancy, primary colonic lymphoma at a single-institute in southern Taiwan.Methods. More than 8953 colon or rectal cancers were reviewed at Chang Gung Medical Center in Kaohsiung from 1989 to 2009, with only 21 cases diagnosed as primary colonic lymphoma. The records of those patients were retrospectively analyzed for age, gender, clinical presentation, location and stage of tumor, operation, adjunctive therapy, survival time and clinical outcome. Cox regression and Kaplan-Meier methods were applied to estimate the prognostic factors for the patient survival time.Results. Twenty-one patients with primary colonic lymphoma were identified which constituted 0.2% of all reported cases of colorectal cancer in our institute. The mean age at diagnosis was 53.9 years (range, 15-86 years), with a male to female ratio of 16:5. The most common tumor site was the cecum, followed by the ascending colon. All patients had B-cell lymphoma. Seventeen patients had undergone an operation. The chemotherapy was applied to 12 patients out of 21. The overall five-year survival rate was 42.9% (N = 21, median survival time: 12.7 months). Prognostic factors including disease stage, operation method (curative vs. palliative), operation timing (elective vs. emergency) and chemotherapy showed a significant influence on the survival time (all p ＜ 0.05).Conclusion. The choice of treatment for primary colorectal lymphoma is multimodality, adequate curative surgical resection in selected patients followed by adjuvant chemotherapy seems to result in a favorable outcome.

Umbilical Metastasis-An Unusual Presentation of Colon Cancer: Report of two Cases and Review of Literature

While the liver and lungs are the most common sites of metastasis from colon cancer, umbilical metastasis is rare. Usually this kind of metastasis becomes evident only when intra-abdominal malignancy is widespread with typical presentation of carcinomatosis demonstrated at laparotomy or on imaging study. We present two cases of colon cancer with umbilical metastasis without extensive dissemination. One patient developed a painful umbilical nodule 1.5 years after an uncomplicated colectomy. Another suffered an indurated umbilical nodule associated with an obstructive sigmoid colon cancer. Pathological analysis of these excised umbilical nodules both demonstrated cutaneous metastasis of colonic origin. Careful surveillance of the periumbilical region is indicated in patients with a colonic malignancy to avoid overlooking possible metastatic implants, in order to provide early curative treatment.