OBJECTIVETo investigate the effect of laparoscopic Roux-en-Y gastric bypass on the short-term immune function of type 2 diabetic patients and to explore the correlation between the immune regulatory effect and blood glucose control.

METHODSClinical data of 28 patients with type 2 diabetes mellitus who underwent laparoscopic Roux-en-Y gastric bypass surgery in our hospital during January 2014 to January 2015 were retrospectively collected. Fasting blood glucose (FBG), glycosylated hemoglobin (HbA1c), immunoglobulin (IgG, IgA, IgM), T lymphocytes CD3(+), killer T cells CD3(+)CD8(+), helper T cells CD3(+)CD4(+), natural killer(NK) cell subsets CD16(+),CD56(+) and B cell subsets CD19(+),CD45(+) levels were detected and compared between before and after surgery. Correlation of immunoglobulin and immune cell subsets with the level of FBG and HbA1c were examined.

RESULTSAfter operation, FBG and HbA1c decreased significantly from (5.78±1.15) mmol/L to (14.21±1.89) mmol/L, and (9.96±0.97)% to (6.87±0.69)%, respectively (all P<0.05), and immunoglobulin (IgG, IgA, IgM) increased significantly [IgG: from(9.41±1.23) g/L to (12.74±1.61) g/L, IgM: from (1.71±0.22) g/L to (2.43±0.39) g/L, IgA: from (1.25±0.26) g/L to (1.97±0.23) g/L, all P<0.05]. Besides, T lymphocytes CD3(+), killer T cells CD3(+)CD8(+) and B cell subsets CD19(+)CD45(+) elevated significantly as well [T lymphocytes CD3(+): from (55.3±3.8)% to (67.6±4.6)%, killer T cells CD3(+)CD8(+): from (15.6±5.3)% to (28.7±4.2)%, B cell subsets CD19(+)CD45(+): from (8.4±3.7)% to (18.1±4.1)%, respectively, all P <0.05]. There was no significant difference in the expression of helper T cells CD3(+)CD4(+) and natural killer cell subsets CD16(+)CD56(+) between pre-operation and post-operation(all P>0.05). Levels of immunoglobulin (IgG, IgA, IgM), CD4(+)/CD8(+) ratio and B cell subsets were negatively correlated with HbA1c and FBG levels (IgG with FBG: r=-0.865, IgA with FBG: r=-0.887, IgM with FBG: r=-0.902, CD4(+)/CD8(+) with FBG: r=-0.956, CD19(+)CD45(+) with FBG: r=-0.834; IgG with HbA1C: r=-0.859, IgA with HbA1C: r=-0.957, IgM with HbA1C: r=-0.843, CD4(+)/CD8(+) with HbA1C: r=-0.912, CD19(+)CD45(+) and HbA1C: r=-0.885, all P<0.05), but the proportion of NK cells was not significantly correlated with HbA1c and FBG (P>0.05).

CONCLUSIONSLaparoscopic Roux-en-Y gastric bypass has good effect on the immune function of patients with type 2 diabetes mellitus. Improvement of immune function is correlated with the control of FBG and HbA1c.

Blood Glucose ; analysis ; Diabetes Mellitus, Type 2 ; immunology ; surgery ; Gastric Bypass ; Glycated Hemoglobin A ; analysis ; Humans ; Immunoglobulins ; blood ; Laparoscopy ; Lymphocyte Subsets ; immunology ; Postoperative Period ; Retrospective Studies

Objective To observe what changes the brown adipose tissue (BAT) of T2DM rat models would have,including morphology,function and specially expressed uncoupling protein (UCP1) after the gastric bypass (Roux-en-Y,RYGB) and to explore the effects of RYGB on BAT of T2DM rat models and its related mechanism in order to provide a theoretical and experimental basis for treatment of T2DM patients with RYGB.Methods SD rats were given a high-fat and high-sugar diet for two weeks,by injecting streptozotocin (STZ) 30 mg/kg intraperitoneally to build models.Blood glucose was measured after 72 h and 1 week by the fast blood glucose meter.The models were built successfully if blood glucose at both times were ≥ 16.7 mmol/L.Feeding environment:individually caged,standard rat feed,natural circadian cycle,indoor temperature (18±2)℃,indoor humidity (50±2)％.50 rats were randomly selected and dividing into four groups according to intervention methods:diabetes operation group (group A,n=10),undergoing RYGB surgery with the whole stomach kept;diabetes sham operation group (group B,n=10),the same anesthesia and incision as the previous RYGB group.The operation mode was anterior gastric wall incision and suture,jejunum transection in corresponding position and in situ anastomosis with the same suture method as group A;diabetes control group (group C,n=10),normally feeding after building models;and the last one was the healthy control group (group D,n=10):no special treatment,adequate water feeding ensured.The rest of rats remained to be used.The body mass (BM),fasting blood glucose (FBG),fasting serum insulin(Fins)before and at the 1st,2nd,4th and 8th week after surgery were measured.The number of transversal ceils was calculated by IPP6.0 image software and the average radius of fat cells was calculated.UCP1 expression was tested with western blot.Results ① The fasting blood glucose,fasting serum insulin level and the body weight of dia betic rats were higher than those of the control group,but the insulin sensitivity index was significantly lower.② HE Staining showed:diabetes operation group (group A) rats,compared with diabetes control group and diabetes sham operation group(group B),had obviously higher brown fat cell counts transversally and average radius,and the difference was statistically significant (P＜0.01).Diabetes operation group (group A) rats had no significant difference from the healthy control group(group D) rats,and the diabetes control group (group C) rats had no significant difference from sham operation group (group B) rats as well.③ Western blot showed that after the gastric bypass surgery,compared with the diabetes sham operation group (group B) and the diabetes control group (group C),UCP1 expression of brown adipose tissue of the diabetes operation group (group A) increased significantly (P＜0.05).The diabetes sham operation group (group B) had no significant difference from the diabetes control group (group C),and the diabetes operation group(Group A) had no significant difference from the healthy control group (Group D) as well (P＞0.05).Conclusion RYGB can reduce the body mass and insulin resistance (IR) of diabetic rats and,at the same time,promote the expression of UCP1 of brown adipose tissue.RYGB might increase the activity of brown adipose tissue by regulating the UCP1 signaling pathway to improve body's insulin resistance.

Objective : To explore the type and control measure of black dot⁃like contaminants in cell cultures. Methods: The growth state of bacteria was investigated under an inverted microscope ; Their morphological characteristics were analyzed by Gram and auramine O staining as well as electron microscopy; 16S rDNA gene sequencing was used to analyze bacterial species ; Drug sensitivity test was used to screen antibiotics against the bacteria;Cryopreserved SH⁃SY5Y cells were resuscitated by cell culture supernatant of RAW264 cells. Results: Inverted microscopic real⁃time observations showed that black dot⁃like substances had two growth states : static and moving.They were negative for Gram staining while positive for auramine O staining. Electron microscopy revealed that they were short rod⁃shaped bacteria with a polar flagellum during moving phase. 16S rDNA gene sequencing showed that these bacteria were phenylobacterium zucineum HLK1. Ceftriaxone , carboxycillin and imipenem were screened by drug sensitivity test to have inhibitory effects on the bacteria , but cell culture experiments showed that they could not remove the bacteria from SH⁃SY5Y cells. Contaminated cells could not be cryopreserved for a long time , but resuscitation with RAW264. 7 cell culture supernatant significantly improved the survival rate of cells. Conclusion The black dot⁃like contaminants in cell cultures are a special type of oligotrophic bacterium with strong viability that can invade the cells and cannot be cleared with antibiotic treatment. RAW264. 7 cell culture supernatant seems contain some substances against bacteria , and resuscitating frozen cells with RAW264. 7 cell culture supernatant may significantly improve the survival rate of cells.