Identification of CD44 as a Cell-Surface Marker for Kit Negative Interstitial Cells of Cajal in Adult Mouse Colon.

Loss of Kit protein expression is proven to influence the plasticity of interstitial cells of Cajal (ICCs) and may contribute to gastrointestinal (GI) dysfunctions. The role and fate of Kit negative ICCs are unclear, and cell-specific markers for the Kit ICCs are unknown. In this study, we treated adult mice with imatinib (a Kit signaling blocker) for 8 or 16 days and investigated whether CD44 is a specific marker for the Kit negative ICCs in the adult mouse colon. We aimed at examining the protein and mRNA level of CD44 and Kit by using Western blot and real-time RT-PCR, respectively. Our results indicated that Kit expression was downregulated for both protein and mRNA levels after imatinib treatment for 8 or 16 days as compared to the vehicle-treated mice. Interestingly, CD44 expression remained unchanged throughout the treatment. Immunostaining on whole-mount preparations for Kit and CD44 showed that CD44 was exclusively co-localized with Kit in the ICCs of the vehicle-treated mouse colon. After imatinib treatment, a number of CD44+/Kit- cells with elaborated processes were observed with an evident decrease of Kit+ cell number within the muscular layers (ICC-IM) and around the myenteric nerve plexus (ICC-MY) as compared to vehicle-treated mice. After discontinuing imatinib for 16 days, Kit+ ICC-MY and ICC-IM were completely co-localized with normalization of CD44 and Kit+ cell numbers. Overall, our results identify CD44 as a cell-specific surface marker for Kit-ICCs and may be useful to understand the role and fate of Kit- ICCs in GI disorders.

[Influence of intensive insulin therapy on insulin resistance of patients with severe burn or trauma].

OBJECTIVE: To discuss the influence of intensive insulin therapy on insulin resistance of patients with severe burn or trauma. METHODS: Sixty patients with severe burn or trauma hospitalized in the Third People's Hospital of Chongqing or Southwest Hospital of the Third Military Medical University from January 2010 to December 2011 were randomly divided into intensive insulin therapy group (IT, treated with intensive insulin therapy to control the blood glucose to the level of 6.0-8.0 mmol/L) and control group (C, treated with routine therapy) according to the paired grouping method, with 30 patients in each group. Before treatment and on post treatment day (PTD) 1, 3, 7, 10, 14, the levels of fasting blood glucose and fasting plasma insulin were determined. Insulin resistance index and ß-cell function index were calculated using homeostasis model assessment. Data were processed with t test, analysis of variance, and LSD test. RESULTS: On PTD 1, 3, 7, 10, levels of fasting blood glucose in group IT [(6.8 ± 1.4), (6.7 ± 1.3), (5.8 ± 1.9), (5.4 ± 1.6) mmol/L] were significantly lower than those of group C [(14.8 ± 4.9), (12.7 ± 3.7), (7.7 ± 1.9), (6.6 ± 1.3) mmol/L, with t values respectively 12.453, 11.386, 5.563, 4.731, P < 0.05 or P < 0.01]. On PTD 3, 7, levels of fasting insulin in group IT [(14 ± 5), (10 ± 3) mU/L] were significantly lower than those of group C [(16 ± 4), (13 ± 4) mU/L, with t values respectively 4.212, 4.364, P values below 0.05]. Levels of fasting blood glucose and fasting insulin in the two groups at each time point were statistically significantly different from those before treatment (with P values below 0.01), except for the level of fasting blood glucose on PTD 3. On PTD 1, 3, 7, 10, levels of insulin resistance index in group IT (1.60 ± 0.80, 1.46 ± 0.70, 0.96 ± 0.21, 0.90 ± 0.23) were significantly lower than those in group C (2.15 ± 1.35, 2.21 ± 1.21, 1.50 ± 0.95, 1.17 ± 0.66, with t values respectively 8.316, 10.607, 7.825, 5.217, P < 0.05 or P < 0.01). Levels of insulin resistance index of patients in the two groups at each time point after treatment were significantly lower than those before treatment (with P values below 0.01). On PTD 1, 3, 7, levels of ß-cell function index in group IT (4.6 ± 2.9, 4.5 ± 3.3, 4.5 ± 3.6) were significantly higher than those in group C (3.4 ± 2.5, 3.6 ± 2.2, 4.2 ± 2.5, with t values respectively 8.243, 7.914, 4.338, P < 0.05 or P < 0.01). Levels of ß-cell function index in group C on PTD 1 and 3 were significantly lower than that before therapy (with P values below 0.05). CONCLUSIONS: Intensive insulin therapy can alleviate insulin resistance of patients with severe burn or trauma.

[Effects of intensive insulin therapy on inflammatory response and prognosis of patients with severe trauma].

OBJECTIVE: To investigate the effects of intensive insulin therapy on inflammatory response and prognosis of patients with severe trauma. METHODS: Eighty severely injured patients were divided into intensive insulin therapy group (n = 40, IT) and routine therapy group (n = 40, RT) in random pair. At the time of admission, a continuous infusion of insulin (2-4 U/h) was pumped into the patients of IT group to maintain blood glucose level at 6-8 mmol/L. Patients in RT group were given routine treatment without administration of insulin. Fever, organ injury, and mortality of patients in 2 groups were recorded. Venous blood was drawn from patients of 2 groups on the morning of post treatment day (PTD) 1, 3, 5, and 7. Values of TNF-alpha, C-reactive protein (CRP), IL-2, and IL-10 in plasma were assayed. RESULTS: High fever appeared in 9 patients in IT group, and WBC exceeded 10.0 x 10(9) for more than 3 days in 17 patients in this group, versus 20 and 29 patients respectively in RT group. Dysfunction of 1 organ appeared in 31 patients in IT group and 30 patients in RT group. Dysfunction of 3 organs appeared in 10 patients in IT group and 19 patients in RT group. Dysfunction of 4 organs appeared in 7 patients in IT group and 12 patients in RT group. In IT group, 4 patients died within 3 post-injury day (PID), and 1 patient died after PID 3 (total case fatality: 12.5%). In RT group, 5 patients died within 3 PID, and 4 patient died after PID 3 (total case fatality: 22.5%). Plasma levels of TNF-alpha and CRP of patients in IT group were significantly lower than those of patients in RT group on PID 3-7 ( P < 0.05 or P < 0.01), while levels of IL-2 and IL-10 of patients in IT group were significantly higher than those of patients in RT group (P < 0.05 or P < 0.01). Plasma levels of TNF-alpha (1.3 +/- 0.6 microg/L) and CRP (55 +/- 16 mg/L) of patients in IT group on PTD 7 were lowered to the trough level, and they were significantly lower than those of patients in RT group (3.0 +/- 0.8 microg/L, 89 +/- 20 mg/L, respectively, P < 0.01). CONCLUSIONS: Intensive insulin therapy can mitigate systemic inflammatory response and improve prognosis of patients with severe trauma.

[Influence of delayed rapid fluid resuscitation on oxygen metabolism in dogs with burn shock].

OBJECTIVE: To investigate the influence of delayed rapid fluid resuscitation on oxygen metabolism in dogs with burn shock. METHODS: Twenty-four mongrel dogs inflicted with 40% TBSA full thickness scald were enrolled in the study and randomly divided into burn control (C), delayed even fluid replacement (E), and delayed rapid fluid replacement (R) groups, with 8 dogs in each group. The changes in oxygen delivery (DO(2)), oxygen consumption (VO(2)), oxygen extraction (O(2)ext) and blood base deficit (BD), and lactate (LA) were determined before scalding and at 2, 6, 8, 12, 24, 36 and 48 post scalding hours (PSHs). RESULTS: The DO(2) in each group was decreased obviously after scalding and was evidently lower than that before injury (P < 0.01), while the O(2)ext value markedly increased compared with that before scalding (P < 0.01). After fluid resuscitation, DO(2) and VO(2) in E and R groups increased, but O(2)ext decreased. The values of DO(2), VO(2) and O(2)ext showed significant differences between R and E groups at 8 PSH (R group vs E group, DO(2): 7.35 +/- 0.21 L.min(-1).m(2) vs 5.32 +/- 0.96 L.min(-1).m(2), P < 0.01; VO(2): 2.02 +/- 0.58 L.min(-1).m(2) vs 1.71 +/- 0.38 L.min(-1).m(2), P < 0.01); The blood BD levels in each group were remarkably lower after scald than that before scald (P < 0.01), and they gradually increased after fluid replacement. The blood BD level in R group at 8 PSH (-6.5 +/- 0.7 mmol/L) was obviously higher than that in E group (-9.3 +/- 1.4 mmol/L, P < 0.01). The blood LA level in each group were evidently higher than that before scald (P < 0.01), and they decreased after fluid replacement. The blood LA level in R group at 8 PSH (2.30 +/- 0.20 mmol/L) was obviously lower than that in E group (2.67 +/- 0.30 mmol/L, P < 0.01) CONCLUSION: Rapid fluid replacement could improve tissue oxygen metabolism, which was beneficial to the correction of tissue oxygen supply when fluid resuscitation was delayed.

[Effect of intensive insulin therapy on serum proinflammatory cytokine levels in patients with severe trauma].

OBJECTIVE: To investigate the effect of intensive insulin therapy on serum levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and C reaction protein (CRP), all of which reflected the inflammatory status in patients with severe trauma. METHODS: Forty patients with severe trauma [injury severity score (ISS)>or=20] were randomly divided into intensive insulin therapy group and control group. Enzyme-linked immunoadsorbent assay (ELISA) method was used to determine the TNF-alpha and IL-6 levels within 72 hours after admission. RESULTS: Serum levels of TNF-alpha, IL-6 and CRP in patients received intensive insulin therapy were significantly lower than those in patients without the therapy (P<0.05 or P<0.01). CONCLUSION: Intensive insulin therapy can attenuate the systemic inflammatory response to trauma. The anti-inflammatory actions of insulin, as well as its glycemia controlling effects, might contribute to the improved outcomes of patients with severe trauma.

[Clinical study of plasma substitute (Gelofusion) on fluid resuscitation in patients with burned shock].

OBJECTIVE: To investigate the effects of plasma substitute(Gelofusion) on fluid resuscitation in patients with burned shock. METHODS: Twenty burn shock patients with total body surface area (TBSA) more than 40% were enrolled for clinical study on the effect of resuscitation with plasma substitute (Gelofusion). The patients were randomly divided into two groups: Gelofusion resuscitation group (n=11) and plasma resuscitation group (n=9). The cardiac output (CO),oxygen delivery (DO2), packed cell volume (PCV), blood and plasma viscosity, lactate(LA) and base deficit (BD) levels were detected at shock stage (postburn from 1 to 48 hours). RESULTS: Two hours after rapid fluid replacement, the levels of CO and DO2 were gradually increased, while the levels of PCV, blood and plasma viscosity, LA and BD were markedly decreased (P<0.05 or P<0.01). After resuscitation, plasma viscosity in Gelofusion resuscitation group were obviously lower than that in plasma resuscitation group within 24 hours postburn (all P<0.05).With an exception of plasma viscosity, there were no significant differences in other parameters between two groups at various intervals (all P>0.05). CONCLUSION: In the resuscitation of burn shock, the clinical effect of Gelofusion treatment is similar to that of plasma treatment. Gelofusion appears to be a fairly good plasma substitute for extensive application on the management of burn shock during the early stage.