Effect of Stem Cells and Gene Transfected Stem Cells Therapy on the Pancreas of Experimentally Induced Type 1 Diabetes

BACKGROUND AND OBJECTIVES: Insulin secretion entirely depends on Ca²⁺ influx and sequestration into endoplasmic reticulum (ER) of β-cells, performed by Sarco-ER Ca²⁺-ATPase 2b (SERCA2b). In diabetes, SERCA2b is decreased in the β-cells leading to impaired intracellular Ca²⁺ homeostasis and insulin secretion. Adipose mesenchymal stem cells (AMSCs) play a potential role in transplantation in animal models. The present study aimed at investigating and comparing the therapeutic effect of non-transfected AMSCs and SERCA2b gene transfected AMSCs on the pancreas of induced diabetes type 1 in rat. METHODS AND RESULTS: 58 adult male albino rats were divided into: Donor group: 22 rats, 2 for isolation, propagation and characterization of AMSCs and SERCA2b transfected AMSCs, in addition 20 for isolated islet calcium level assessment. Group I (Control Group): 6 rats, Group II (Diabetic Group): 10 rats, 50 mg streptozotocin (STZ) were injected intraperitoneal (IP), Group III (AMSCs Group): 10 rats, 1×10⁶ AMSCs were injected intravenous and Group IV (SERCA2b transfected AMSCs Group): 10 rats, 1×10⁶SERCA2b transfected AMSCs were injected as in group III. Groups I, II, III and IV were sacrified 3 weeks following confirmation of diabetes. Serological, histological, morphometric studies and quantitative polymerase chain reaction (qPCR) were performed. Nuclear, cytoplasmic degenerative and extensive fibrotic changes were detected in the islets of group II that regressed in groups III and IV. Isolated islet calcium, blood glucose, plasma insulin and qPCR were confirmative. CONCLUSIONS: AMSCs and SERCA2b gene transfected AMSCs therapy proved definite therapeutic effect, more obvious in response to SERCA2b gene transfected AMSCs.

Efficacy and safety of intraoperative radio frequency ablation in management of hepatocellular carcinoma

The present study is a prospective one involving 15 consecutive patients with hepatocellular carcinoma [HCC] from January 2003 to December 2005. Clinical and laboratory examinations, abdominal ultrasonography and spiral CT scanning were performed. All patients were subjected to intraoperative radiofrequency ablation of HCC after confirmation by core biopsy. Enhanced dynamic CT was done at 1 month postoperative and every 3 months during follow-up period. Males were 12 with age of 41-69 year [53.2 +/- 7.4]. Underlying cirrhosis due to viral hepatitis was in all, and HCV was the most common [75%], and patients [85%] were categorized in Child B classification. Serum alpha feto protein was above 400 ng/ml in 60%. 12 patients had unifocal HCC [nodules mean diameter of less than 3 cm in 6, 3 to 5 cm in 4, and more than 5 cm in 2], and 3 patients had multifocal HCC [nodules mean number of less than 3 cm in 2 and 3 to 5 cm in 1]. The mean number of RF application session to achieve complete necrosis in patients with were 1.52 [unifocal] and 2.49 [multifocal HCC]. The mean hospital stay was 14.9 days, with neither mortality nor major complications, but Minor complications in 30%. After one month, complete ablation of HCC nodules was achieved in 10 patients with reduction of alpha fetoprotein in 11 patients. 2/5 with partial ablation were multifocal nodules. Total mortality during follow-up in late post-operative period [1 year] was 6 out of 15 patients. 5 of them are the patients with partial ablation. 3 patients out of 9 patients developed evidence of local recurrence detected by enhanced dynamic CT and raising of serum alpha fetoprotein

Thyroxine as a stimulator of liver regeneration after partial hepatectomy: an experimental animal study

Twenty six male albino rats were divided into a control group [6rats] [G] exposed to Sham operation and two experimental groups [Gl and G2] of 10 rats each. 2/3 partial hepatectomy [PH] was performed in Gl and T4 was administered orally in a dose of 4mg/kg 10 days prior to PH in G2. Animals were sacrificed 24 hours following PH and Sham operation. Liver sections were immunohistochemically stained for anti-cyclin Dl and anti-VEGF antibodies. Positive reaction was in the nuclei of hepatocytes and in lining of hepatic blood sinusoids respectively. The morphometric determination of the count of cyclin Dl positive nuclei and area% VEGF positive lining of the blood sinusoids was performed in the different groups. Multiple cyclin Dl positive nuclei were detected in liver sections of rats receiving thyroxine prior to PH confirmed by significant increase in the mean count of immuno-positive nuclei as compared to G and Gl. VEGF positive immunoreaction was in the lining of multiple hepatic blood sinusoids on thyroxine therapy proved morphometrically by a significant increase in mean area% in G2 versus Gl. The results showed a stimulating effect of thyroxine on liver regeneration following 2/3 partial hepatectomy evidenced by increased Cyclin D and sinusoidal endothelial VEGF expression

Soluble P-and E- selectins and ultrastructural evaluation of canine liver after reperfusion with methylprednisolone and prostaglandin E 1

Ischemia reperfusion injury [IRI] is a multifactorial process that may be the main underlying factor in critical phases faced during and after liver transplantation. This work evaluates the effect of methylprednisolone [MP] and prostaglandin El [PGE1] on IRI of the liver of dogs at the ultrastructural level together with the assessment of soluble P-and E- selectin levels. Three groups of dogs [9 each] were subjected to 60 min ischemia followed by 30 min reperfusion with the appropriate solution according to the group. Group I, the control untreated group was flushed with Ringer's solution, group II was administered 10 mg/kg MP 24 hours before the procedure and with induction of anaesthesia, and group III was flushed with PGE1 in Ringer's solution at a rate of 0.02 microg/kg/min. Liver specimens were collected before ischemia, after ischemia and after reperfusion and were processed for the preparation of ultrathin sections for electron microscopic examination. Corresponding venous blood samples were harvested, centrifuged and serum was processed for the estimation of soluble P-and E- selectins by enzyme immunoassay, together with alanine and aspartate aminotransferases. Electron microscopic [EM] examination of liver ultrathin sections revealed that the morphological structure of hepatocytes and endothelial lining of hepatic sinusoids were well preserved in the group treated with PGE1. Hepatic ultrastructure was much altered in MP treated group showing necrotic and degenerative changes. The control untreated group disclosed bleb formation of hepatocytic membrane with increased leukocyte infiltration. Soluble P-and E- selectin levels were significantly elevated in the control group, near to pre-ischemic level in PGE1 group and showed a persistent elevation in MP group. Serum transaminases [AST and ALT] were significantly elevated in both control and MP groups as compared to their corresponding pre-ischemic levels. Yet, in PGE1 group, their values were comparable to the pre-ischemic ones. This work confirms the hepatoprotective effect of PGE1 in IR injury. The PGE1 impact on preserving the subcellular structure of hepatic sinusoids is crucial and may be mainly attributed to its biological properties. We presume that P-and E- selectins are greatly implicated in the mechanism of IR and may be an important therapeutic target by specific monoclonal antibodies