[The role of mesenteric lymph in pathogenesis of systemic inflammatory response syndrome and systemic complications following severe acute pancreatitis in rats].

OBJECTIVE: To observe the effect of mesenteric lymph duct ligation (MLDL) on systemic inflammatory response syndrome (SIRS) and systemic complications in severe acute pancreatitis (SAP) in rats. METHODS: Twenty-four Sprague-Dawley (SD) male rats were randomly divided into sham operation group, SAP group, and SAP+MLDL group, with 8 rats in each group. Sham operation group underwent laparotomy only. SAP complicated with multiple organ injury model was reproduced by retrograde injection of sodium taurocholate into the pancreatic duct in SD rats. In SAP+MLDL group MLDL was performed before reproduction of SAP. Rats were sacrificed at 24 hours after operation. The terminal ileum were collected for examination under light microscope. The samples of pancreas, lung and liver were collected for measuring the level of myeloperoxidase (MPO). The mesenteric lymph nodes were harvested for standard bacteriologic culture. The blood levels of diamine oxidase (DAO), D-lactate, endotoxin, tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) were also measured. RESULTS: The blood levels of DAO [(0.64 + or - 0.17) kU/L vs. (0.37 + or - 0.07) kU/L], D-lactate [(8.16 + or - 1.79) ng/L vs. (3.24 + or - 1.00) ng/L], endotoxin [(0.068 + or - 0.019) kEU/L vs. (0.033 + or - 0.009) kEU/L], TNF-alpha [(65.21 + or - 13.38) ng/L vs. (22.16 + or - 5.04) ng/L] and IL-6 [(7.95 + or - 1.83) ng/L vs. (4.26 + or - 1.23) ng/L] in SAP group were increased significantly comparing with those in sham operation group (all P<0.01). The contents of pulmonary tissue [(9.07 + or - 2.52) U/g vs. (4.38 + or - 1.29) U/g], pancreatic [(5.14 + or - 1.24) U/g vs. (2.88 + or - 0.75) U/g] and liver [(6.36 + or - 1.63) U/g vs. (3.19 + or - 0.96) U/g] MPO were increased significantly in SAP group comparing with those in sham operation group (all P<0.01). The rate of bacterial translocation to mesenteric lymph nodes in SAP group was significantly higher than that of the sham operation group (75.0% vs. 0, P< 0.01). Compared to SAP group, the contents of DAO [(0.50 + or - 0.13) kU/L], D-lactate [(6.23 + or - 1.25) ng/L] , endotoxin [(0.048 + or - 0.014) kEU/L], TNF-alpha [(48.50 + or - 13.23) ng/L] and IL-6 [(6.06 + or - 1.64) ng/L] were significantly decreased in SAP+MLDL group (P<0.05 or P<0.01). The levels of pulmonary [(6.58 + or - 1.96) U/g], pancreatic [(4.01 + or - 1.05) U/g] and liver [(4.64 + or - 1.34) U/g] MPO, as well as the rate of bacterial translocation to mesenteric lymph nodes (12.5%) were significantly lower in SAP+MLDL group than those in SAP group (all P<0.05). CONCLUSION: MLDL decreases bacteria/ endotoxin translocation and reduces the systemic inflammatory response by interrupting the lymph flow from the injured gut into the bloodstream. This procedure may protect the small intestine mucosal barrier function and other organs by reducing the neutrophil aggregation and the release of cytokines.

[Effect of blood glucose control after small intestine exclusion surgery in Goto-Kakizaki rat with type 2 diabetes mellitus].

OBJECTIVE: To evaluate the effect of diabetic control after small intestine exclusion surgery in Goto-Kakizaki (GK) rat with type 2 diabetes mellitus. METHODS: GK type 2 diabetic rats underwent duodenal bypass (Group A, n = 8) creating a shortcut for ingested nutrients with bypassing duodenum alone, or duodenal-jejunal bypass (Group B, n = 8), a stomach-preserving RYGB excluding duodenum and proximal jejunum, or duodenum and total jejunum exclusion (Group C, n = 8), or sub-total small intestine exclusion (Group D, n = 8) creating a shortcut for ingested nutrients with bypassing duodenum, jejunum and sub-total ileum. Controls were pair-fed (PF) sham-operated and untreated GK rats (Group SO, n = 8). Rats were observed for 24 weeks after surgery. Fasting blood glucose (FBG) level was determined on a Surestep plus blood glucose meter (Lifescan) at 0, 1, 3, 6, 12, 24 week. Hemoglobin A1c (HbA1c) level was measured at 0, 12, 24 week on VARIANT Hemoglobin A1c Program (Bio-Ray) with high performance liquid chromatography (HPLC) method. RESULTS: The fasting blood glucose and HbA1c levels significantly decreased after surgery in all the operative groups through the entire follow-up period [Group B (FBG before surgery and 1 week after: (162 +/- 44) mg/dl vs (80 +/- 19) mg/dl; HbAlc before surgery and 12 week after: (8.2 +/- 2.2)% vs (5.1 +/- 1.5)%, P < 0.05 or P < 0.01]; while FBG had no differences before 12 week and significantly increased at 12 week (164 +/- 44) mg/dl and 24 week (180 +/- 42) mg/dl in group SO [preoperative (145 +/- 35) mg/dl, P < 0.01]. As compared with sham-operated rats, all the operative groups showed reduced blood-glucose and HbA1c levels through the entire follow-up period (P < 0.05 or P < 0.01). The FBG and HbA1c levels in Group B significantly decreased versus Group A [24 week (FBG: (82 +/- 21) mg/dl vs (111 +/- 27) mg/dl; HbA1c: (3.9 +/- 0.9)% vs (5.4 +/- 1.2)%, P < 0.05], indicating that duodenal-jejunal bypass had markedly a better effect of diabetic control. But Groups B, C and D were similar to one another (P > 0.05), suggesting that a potential role of proximal gut in the pathogenesis of disease. CONCLUSION: In spontaneous type 2 diabetes mellitus rats, all the small intestinal exclusion including proximal gut are effective in glucose control. In terms of intestinal nutrient absorption, duodenal-jejunal bypass may be an ideal surgery for clinical diabetic control. These findings suggest a potential role of proximal intestine in pathogenesis of type 2 diabetes mellitus.

[Effect of diabetes control after small intestine exclusion surgery in Goto-Kakizaki rat with non-obese type 2 diabetes mellitus].

OBJECTIVE: To evaluate the effect and possible mechanisms of diabetes control after small intestine exclusion surgery in Goto-Kakizaki (GK) rat with non-obese type 2 diabetes mellitus. METHODS: Forty GK rats with non-obese type 2 diabetes mellitus underwent duodenal bypass (Group A, n = 8), which creates a shortcut for ingested nutrients bypassing duodenum alone; duodenal-jejunal bypass (Group B, n = 8), a stomach-preserving RYGB that excludes the duodenum and proximal jejunum; duodenum and total jejunum exclusion (Group C, n = 8); sub-total small intestine exclusion (Group D, n = 8), which creates a shortcut for ingested nutrients bypassing duodenum, jejunum and sub-total ileum; controls were pair-fed (PF) sham-operated and untreated GK rats (Group SO, n = 8). The rats were observed for 24 weeks after surgery. Body weight, food intake and fasting blood glucose level were tested at 0, 1, 3, 6, 12, 24 weeks after the operation in those groups. The concentrations of insulin and glucagon-like peptide-1 (GLP-1) concentrations were measured before (baseline) and then 30, 60, 120, and 180 minutes after OGTT at 0, 12, 24 weeks after the operation. RESULTS: Mean operating time of all groups was similar. The mean body weight and food intake decreased significantly at 1 week after surgery (P < 0.01) and with no differences among the groups. Fasting blood glucose level decreased significantly after surgery in all the operation groups through the entire follow-up period (P < 0.05). No significant changes in fasting blood glucose level in SO group was found in 12 weeks after the operation, and it increased at 12 and 24 weeks after. Fasting blood glucose levels in group B decreased significantly compared with group A (P < 0.05), but no difference was found among group B, C and D (P > 0.05). Oral glucose-stimulated peak (30 min) levels of blood insulin and GLP-1 increased markedly in operation groups (A, B, C and D) after surgery (P < 0.01). Compared with group A, peak levels of blood insulin and GLP-1 in group B were strikingly higher (P < 0.05), but no difference was found when compared with group C or D (P > 0.05). CONCLUSIONS: In spontaneously non-obese type 2 diabetes mellitus rats, small intestinal exclusion including proximal gut is effective in terms of glucose control and has no direct relation with body weight and food intake loss. Restoration of the first-phase insulin secretion and high secretion of GLP-1 in type 2 diabetic subjects after gastrointestinal bypass surgery seem to be helpful to diabetes control. Taking intestinal nutrient absorption into consideration, duodenal-jejunal bypass may be a better surgery for diabetes control.

[Effects of Xuesaitong injection on coagulation function of gastric cancer patients undergoing laparoscopic surgery: a randomized controlled trial].

OBJECTIVE: To study the effects of Xuesaitong Injection, an extract from a Chinese herbal medicine, on coagulation function of gastric cancer patients undergoing laparoscopic surgery. METHODS: Fifty patients undergoing laparoscopic surgery for gastric cancer were randomly divided into control group and Xuesaitong group, with 25 patients in each group. Conventional anti-infection and transfusion treatment was given to the patients in the control group, and intravenous drip infusion of 400 mg Xuesaitong Injection added into 250 mL 5% glucose injection was administered once daily to the patients in the Xuesaitong group for 6 days from the first day after laparoscopic operation besides the conventional anti-infection and transfusion treatment. The values of prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) and international normalized ratio (INR) were detected by using blood coagulation method, and the values of D-dimer (D-D) before operation, instantly and 3, 5, 7 days after operation were detected by using enzyme-linked immunosorbent assay in the two groups. Furthermore, the development of deep vein thrombosis on the day 7 after operation was observed by vein angiography. RESULTS: There were no significant differences in the values of PT, APTT, INR, FIB and D-D between the two groups before operation, and there were no significant changes in APTT and INR after operation in the two groups. The values of PT decreased in the two groups on day 3 after operation, while there was no significant difference between the two groups. The value of PT in the control group decreased gradually with the extension of postoperation time (P < 0.05), while the PT value in the Xuesaitong group did not change significantly, and there was significant difference between the two groups (P < 0.05). The values of FIB and D-D increased instantly after operation in the two groups as compared with those before operation (P < 0.05), while there was no significant difference between the two groups. With the extension of postoperation time, the values of FIB and D-D increased gradually in the control group (P < 0.05); the D-D value on day 3 after operation increased, but the D-D values on day 5 and day 7 after operation decreased gradually in the Xuesaitong group (P < 0.05). The occurrence rate of deep vein thrombosis on day 7 after operation in the Xuesaitong group was lower than that in the control group. CONCLUSION: Laparoscopic surgery for gastric cancer may cause high agglutination condition, which will increase the risk of deep vein thrombosis, and Xuesaitong Injection can improve this state.

Protective effect of transplanted bone marrow-derived mesenchymal stem cells on pancreatitis-associated lung injury in rats.

Severe acute pancreatitis (SAP) is initiated by the premature activation of digestive enzymes within the pancreatic acinar cells, leading to self-digestion and inflammatory responses in pancreatic ductal cells, thus giving rise to systemic inflammatory response syndrome (SIRS). The most common and serious SIRS is pancreatitis-associated lung injury, and inflammatory mediators play an important role in its pathogenesis. Bone marrow-derived mesenchymal stem cells (MSCs) are differentiated into alveolar endothelial cells to replace the damaged alveolar endothelial cells and inhibit inflammatory response in the injured lung tissues. In this study, we aimed to investigate the therapeutic effect of bone marrow-derived MSCs in rats with pancreatitis-associated lung injury. Experimental SAP was induced by a retrograde injection of 5% sodium taurocholate into the biliopancreatic duct of 75 male Sprague-Dawley rats, which were divided into the SAP group (n=25), the MSC group (n=25) and the sham-operated group (n=25) to explore the pathology and function of lung tissues and the regulation of inflammatory mediators. Pulmonary edema was estimated by measuring water content in the lung tissues. Pulmonary myeloperoxidase (MPO) activity was detected using spectrophotometry. Serum amylase was detected using the Automatic Biochemistry Analyzer. Tumor necrosis factor-α (TNF-α) and substance P (SP) mRNA levels were determined by quantitative reverse transcriptase-polymerase chain reaction. Our results showed that serum amylase activity was significantly decreased in the MSC group compared to the SAP group. Pulmonary edema was significantly diminished (p<0.05) in the MSC group compared to the SAP group. Typical acute lung injury was observed in the SAP group, and the pathological changes were mild in the MSC group. The expression of TNF-α and SP mRNA in lung tissue was diminished in the MSC group compared to the SAP group. In conclusion, MSC transplantation attenuates pulmonary edema and inflammation, and reduces the mRNA expression of TNF-α and SP in pancreatitis-associated lung injury.

[Comparative effects of Roux-en-Y gastric bypass procedures preserving different gastric volume on blood glucose in Goto-Kakizaki rats].

OBJECTIVE: To compare the effects of Roux-en-Y gastric bypass (RYGBP)procedures preserving different gastric volume on blood glucose of rats with non-obese type 2 diabetes. METHODS: A total of 36 Goto-Kakizaki rats randomly underwent one of the following procedures: gastric bypass with different types of anastomosis including the Roux-en-Y of total stomach excision(n=12), the Roux-en-Y of partial stomach excision(n=12) and the Roux-en-Y of stomach preservation(n=12). Rats were observed for 24 weeks after surgery. Body weight, food intake and fasting blood glucose level were tested at 0(preoperative), 1, 3, 6, 12, 24 weeks. Hemoglobin A1c(HbA1c) level was measured at 0, 12, 24 weeks and glucose tolerance test (OGTT) was performed in conscious rats before (baseline) and then 30, 60, 120, and 180 minutes. Change of blood glucose over time was depicted. Area under curve(AUC) of glucose tolerance were calculated. RESULTS: Compared with preoperative levels, the weight and food intake of all the rats were significantly decreased at 1 week after surgery(P<0.01). At 3 weeks after operation, the weight and food intake were significantly increased compared with 1 week after operation in the Roux-en-Y of partial stomach excision and the Roux-en-Y of stomach retention(P<0.01). In the Roux-en-Y of total stomach excision, the weight and food intake were significantly lower compared with other two groups(P<0.05). At 24 weeks after operation, the levels of fasting blood glucose were (7.3 ± 1.5), (7.5 ± 2.0) and (8.3 ± 1.3) mmol/L, which were lower than the preoperative levels [(13.2 ± 1.6), (13.6 ± 2.5) and (12.9 ± 2.0) mmol/L, P<0.01] in the three groups. There were no significant differences among the three groups(P>0.05). At 24 weeks after operation, the HbA1c levels were(6.3 ± 1.3)%, (6.4 ± 2.0)% and (7.0 ± 1.3)%, which were lower than the preoperative level[(10.2 ± 2.6)%, (9.6 ± 2.5) and (9.9 ± 2.0)%, P<0.01]. There were no significant differences among the three groups(P>0.05). The trend of the glucose tolerance test and AUC were similar in the three groups after operation. CONCLUSION: Roux-en-Y gastric bypass in non-obese diabetic rats is effective in terms of glucose control and the efficacy of gastric bypass has no obvious association with the stomach volume.

Role of bone marrow-derived mesenchymal stem cells in a rat model of severe acute pancreatitis.

AIM: To investigate the role and potential mechanisms of bone marrow mesenchymal stem cells (MSCs) in severe acute peritonitis (SAP). METHODS: Pancreatic acinar cells from Sprague Dawley rats were randomly divided into three groups: non-sodium deoxycholate (SDOC) group (non-SODC group), SDOC group, and a MSCs intervention group (i.e., a co-culture system of MSCs and pancreatic acinar cells + SDOC). The cell survival rate, the concentration of malonaldehyde (MDA), the density of superoxide dismutase (SOD), serum amylase (AMS) secretion rate and lactate dehydrogenase (LDH) leakage rate were detected at various time points. In a separate study, Sprague Dawley rats were randomly divided into either an SAP group or an SAP + MSCs group. Serum AMS, MDA and SOD, interleukin (IL)-6, IL-10, and tumor necrosis factor (TNF)-α levels, intestinal mucosa injury scores and proliferating cells of small intestinal mucosa were measured at various time points after injecting either MSCs or saline into rats. In both studies, the protective effect of MSCs was evaluated. RESULTS: In vitro, The cell survival rate of pancreatic acinar cells and the density of SOD were significantly reduced, and the concentration of MDA, AMS secretion rate and LDH leakage rate were significantly increased in the SDOC group compared with the MSCs intervention group and the Non-SDOC group at each time point. In vivo, Serum AMS, IL-6, TNF-α and MAD level in the SAP + MSCs group were lower than the SAP group; however serum IL-10 level was higher than the SAP group. Serum SOD level was higher than the SAP group at each time point, whereas a significant between-group difference in SOD level was only noted after 24 h. Intestinal mucosa injury scores was significantly reduced and the proliferating cells of small intestinal mucosa became obvious after injecting MSCs. CONCLUSION: MSCs can effectively relieve injury to pancreatic acinar cells and small intestinal epithelium, promote the proliferation of enteric epithelium and repair of the mucosa, attenuate systemic inflammation in rats with SAP.

[Protective effect of N-acetylcysteine on the intestinal barrier dysfunction after radiation injury in rats].

OBJECTIVE: To evaluate the protective effect of N-acetylcysteine (NAC) on the intestinal barrier dysfunction in rats after extensive abdominal radiation with X ray. METHODS: Twenty-four Spraque-Dawley male rats were divided into normal control group (n=8), radiation group (n=8), and radiation+NAC group (300 mg/kg) (n=8). Radiation injury was induced by X ray with a single dose of 10 Gy. NAC was administered from 4 days before irradiation to 3 days after radiation. Three days after radiation, all the rats were euthanized. The terminal ileum was collected for crypt survival assay and ileal villi count. The tissue samples from mesenteric lymph nodes (MLN), spleen, and liver were harvested under sterile conditions for microbiological analysis and ileum samples were harvested for biochemical analysis. The blood levels of D-lactate, endotoxin and diamine oxidase (DAO) and the ileum samples levels of nitric oxide(NO) were also measured. RESULTS: Rats in radiation+NAC group had a higher survival rate of intestinal crypt [(76.84+/-4.82)% vs (49.64+/-5.48)%, P<0.01], higher intestinal villus count [(8.56+/-0.68)/mm vs (4.02+/-0.54)/mm, P<0.01], lower NO concentration [(0.48+/-0.12) mumol/g vs (0.88+/-0.16) mumol/g, P<0.01], lower levels of D-lactate, endotoxin and DAO (P<0.05 or P<0.01), and significantly decreased enteric bacteria cultured from mesenteric lymph nodes and other tissues as compared with the radiation group (P<0.05 or P<0.01). CONCLUSION: NAC protects the small intestine from radiation-induced injury maybe through the inhibition of NO in rats.

[Evaluation of endoscopy-assisted laparoscopic surgery in the treatment of early colon cancer].

OBJECTIVE: To evaluate the endoscopy-assisted laparoscopic surgery in the treatment of early colon carcinoma. METHODS: The data of 55 early colon cancer patients, including 30 male, 25 female with mean age of 54 years(ranged 42 to 68), undergone endoscopy-assisted laparoscopic surgery at the colon were reviewed retrospectively. RESULTS: From March 2002 to December 2007, 55 early colon cancer patients were treated with endoscopy-assisted laparoscopic surgery in our institute. In 53 cases, a laparoscopic and endoscopic cooperative bowel segment resection was performed at first. Of these 53 patients, 11 cases then received laparoscopic and endoscopic cooperative radical anatomical resection according to the result of frozen section. Two cases were transferred to open surgery because of small intestinal inflation after endoscopic location. The mean operative time of cooperation was 90 min (55-240 min), and the mean blood loss was 50 ml(10-200 ml). In 51 cases(92.7%), the time for flatus passage was 2 to 3 days. The mean postoperative hospital stay was 5 d(2-15 d). No postoperative complications were found. Follow-up data were obtained by clinical examination and personal communication via telephone. The median follow-up was 42 months(ranged 3-72). Most of the patients were alive except one case died of myocardial infarction during the follow-up period. None of the patients with early colon cancer treated by the cooperative surgery had relapse or metastasis. Two cases of T1N1Mx underwent adjuvant chemotherapy. CONCLUSIONS: Endoscopy-assisted laparoscopic surgery offers a minimal-invasive and safe therapeutic approach for early colon cancer. The early colon cancer may be a good indication for endoscopy-assisted laparoscopic surgery when the endoscopic mucosal resection is inadequate.

Safety and efficacy of Qingre Buyi Decoction in the treatment of acute radiation proctitis: a prospective, randomized and controlled trial.

OBJECTIVE: To investigate the efficiency, safety, and possible mechanisms of Qingre Buyi Decoction (QBD) in the treatment of acute radiation proctitis (ARP). METHODS: This study was a single center, prospective, single blind, randomized, and placebo-controlled clinical trial. A total of 60 patients with ARP was equally and randomly distributed into the control group (conventional treatment) and the combination group (conventional treatment plus QBD). The changes of main Chinese medicine clinical symptoms and signs, including stomachache, diarrhea, mucous or bloody stool before and after treatment, and their adverse reactions were observed after the two-week treatment. Also, D-lactate and diamine oxidase (DAO) levels, hepatic and renal function were measured. Cure rates, effective rates, and recurrence rates were compared between the two groups. RESULTS: The blood levels of both DAO and D-lactate were significantly decreased in the combination group as compared with those in the control group (P<0.05 or P<0.01). All main clinical symptoms and signs were alleviated more significantly in the combination group (P<0.01). The main symptom scores also were significantly decreased after treatment in the control group (P<0.01), except those for mucous or bloody stool (P>0.05). Compared to the control group, the improvements of stomachache, diarrhea, defecation dysfunction, and stool blood in the combination group were significantly better (P<0.05 or P<0.01). For the combination group, the curative rate, effective rate, and recurrence rate was 76.67%, 16.67%, and 6.67%, respectively. On the other hand, for the control group, the rate was 53.33%, 16.67%, and 30.00%, respectively. The total curative effect was significantly better in the combination group than in the control group (P<0.05). However, the recurrence rate was similar between the two groups (P>0.05). The hepatic and renal function remained normal in both groups (P>0.05). In addition, no severe adverse event was found in both groups. CONCLUSIONS: Addition of QBD to the conventional treatment can effectively alleviate the damage of intestinal mucosal barrier function and improve all main clinical symptoms and signs of the ARP. The combination of conventional treatment with Chinese herbal medicine QBD is effective and safe for ARP.

[Effect of different position and CO(2) pneumoperitoneum on deep vein hemodynamics of lower limb].

OBJECTIVE: To study the effect of laparoscopic surgery position and CO(2) pneumoperitoneum on deep vein hemodynamics of lower limb. METHODS: Color Doppler ultrasound was adopted to evaluate the diameter, blood velocity and the amount of the blood flow in the lower extremity of 37 gastric cancer patients undergone laparoscopic surgery from November 2007 to August 2008 in our hospital. Three kinds of position(prostration, head-down tilt position and feet-down tilt position) before the establishment of pneumoperitoneum and three different pressure of CO(2) pneumoperitoneum( 8 mm Hg, 12 mm Hg, 16 mm Hg ) after the establishment of pneumoperitoneum were applied during laparoscopic surgery. RESULTS: Compared with the stage of prostration before the pneumoperitoneum, the femoral vein diameter decreased (P<0.05), the velocity and the amount of blood flow increased (P<0.01, P<0.05) in head-down tilt position; the femoral vein diameter increased(P<0.01), the velocity and the amount of blood flow decreased(P<0.01, P<0.05) in feet-down tilt position or the establishment of pneumoperitoneum. Furthermore, with the increase of pneumoperitoneum pressure, the femoral vein diameter increased, the velocity and the amount of blood flow decreased(P<0.05). CONCLUSIONS: The head-down tilt position is conducive for the blood to recirculate, which will degrade the risk of deep vein thrombosis after the laparoscopy. The feet-down tilt position and CO(2) pneumoperitoneum can interfere the recirculation of blood, and increase the risk of deep vein thrombosis after the laparoscopy.