[The Pharmacokinetics, Pharmacodynamics and Bioequivalence of Insulin Aspart Produced by the United Laboratories Evaluated by Euglycemic Clamp Study].

OBJECTIVE: To investigate the pharmacokinetics (PK) and pharmacodynamics (PD) of a rapid-acting insulin analog-insulin aspart (the tested formulation) which was manufactured by The United Laboratories and to evaluate the bioequiavailability to the reference formulation (NovoRapid ®) produced by Novo Nordisk in Chinese healthy volunteers. METHODS: A total of 24 male healthy volunteers were recruited from February to April 2016 to participant in this before-after, single dose, and randomized crossover study. And the experimental observation was conducted on 2 test days respectively with a between-period from 7 to10 d. According to a random number table, the volunteers were divided into group A or B, group A was administrated with tested insulin aspart (IAsp) for the first time and reference NovoRapid ® for the second time and group B had the revered order differed from group A. The PK/PD of these insulin analogs were estimated by euglycemic clamp study. RESULTS: The relative biological effectiveness (reflecting gloucose-lowing effect) and bioavailability on behalf of plasma-drug concentration were 98.3±18.8% and 97.3%±8.3% respectively. For PK parameters, the 90% confidence interval ( CI) of peak plasma insulin concentration ( C max) and area under the curve of insulin aspart concentration from 0 to 10 hours ( AUC IAsp, 0-10 h) of IAsp were 88.8%-106% (equivalent range 70%-143%) and 94.0%-100% (equivalent range 80%-125%) respectively; for PD parameters, the 90% CI of the maximum glucose infusion rate ( GIR max) and AUC GIR, 0-10 h were 95.5%-113% (equivalent range 70%-143%) and 89.9%-104% (equivalent range 80%-125%) respectively, which indicated that IAsp and NovoRapid® was bioequivalent. One of the subjects discovered hyperuricemia without clinical symptoms and the rest had no clinically significant abnormalities in the safety indexes before and after the tests. No hypoglycemic events, allergic reactions, or local injection adverse reaction occurred in this trial. CONCLUSION: The tested IAsp has comparable relative bioavailability to the reference NovoRapid ®.

[Quality Assessment of Euglycemic Glucose Clamp Test].

OBJECTIVE: To explorethe quality of euglycemic glucose clamptest performed in the West China Hospital from 2014 to 2017 and to evaluate whether the quality control indexes are suitable for the quality assessment of the clamp test. METHODS: The data collected from 80 euglycemic glucose clamp tests performed between 2014 and2017 were divided into 4 groups according to the coefficient of variation of the blood glucose concentrations (CVBG): group A (CVBG≤4.5%), group B (4.5% < CVBG≤5.0%), group C (5.0% < CVBG≤5.5%) and group D(CVBG > 5.5%).The differences in percentage of glucose excursion from target range (GEFTR), the duration of GEFTR, the area under curve (AUC) of GEFTR, the mean value of excursion from target glucose (GEFT) and the AUC of GEFT were calculated and compared. RESULTS: In group A, the mean value of CVBG was 3.75%. In group B, the mean value of CVBG was 4.76%. In group C, the mean value of CVBG was 5.28%. The median value of CVBG in group D was 6.07%. The percentage of GEFTR, the duration of GEFTR, the AUC of GEFTR, the mean value of GEFT and the AUC of GEFT in group A were all less than those of other groups (P < 0.05).For the same indexes, there were no significant differences between group B and C, while they were higher in group D compared with the other three groups. CVBG was positively correlated with other quality control indexes (correlation coefficient r was 0.770-0.805). Based on the cut-off point 5% of CVBG, the cut-off points of the percentage of GEFTR, the duration of GEFTR, the AUC of GEFTR, the mean value of GEFT and the AUC of GEFT were 5.8%, 14.6 min, 22.82 mg/dL×min, 3.23 mg/dL, 216.25 mg/dL×min/h respectively, with the sensitivity range from 79.3% to 100% and the specificity range from 74.5% to 89.7%.Combined with these indexes, 8.11% of euglycemic clamps were found to havepoor quality in group A, while 66.67% of euglycemic clamps showed acceptable quality in group C. CONCLUSIONS: The investigators should provide an estimation of the quality of the clamps when reporting the results of the insulin analogues' PK/PD characteristics using euglycemic clamps. CVBG less than 4.5% indicates a good quality, and the above-mentioned quality control indexes especially the AUC of GEFT(cut-off point: 216.25 mg·/dL×min/h) should be evaluated when CVBG is more than 4.5%.False high quality and false low quality euglycemic clamps will be detected and a more precise estimation of quality assessment should be made by the combination of these indexes.

Multidrug resistance and tumor-initiating capacity of oral cancer stem cells.

PURPOSE: Recent studies in several tumors showed that presence of cancer stem like side population (SP) cells are responsible for chemotherapeutic drugs resistance and tumor relapse. In our present study, we have analyzed the role of SP cells in oral squamous cell carcinoma cell (OSCC) line OSCC-77. METHODS: The oral cancer cell line OSCC-77 was analyzed for the presence of SP cells by FACS using Hoechst 33342 dye exclusion method. Further the FACS-sorted SP and non-SP cells were subjected to drug resistance and sphere formation assays. RESULTS: We identified that the presence of SP cells in OSCC-77 cell line was 3.4%, which was reduced to 0.6% in the presence of verapamil, an inhibitor of ABC transporter. Furthermore, we showed that these SP cells were highly drug-resistant, had increased survival and were highly potent for self-renewal. Also, the clone formation efficiency of SP cells was significantly higher compared to non-SP cells (p<0.01). CONCLUSION: Our data suggest that cancer stem-like SP cells of OSCC-77 cell line contribute to multidrug resistance and are highly involved in tumor relapse. However, further characterization of SP cells at gene expression level and their signaling pathways might provide new insights into the development of novel anticancer drugs.

[Association of nt3434A→G Mutation in Mitochondrial DNA ND1 Gene with Diabetes Mellitus].

OBJECTIVES: To study the association of nt3434 A→G mutation in mitochondrial DNA NADH dehydrogenase 1 subunit (ND1) gene with diabetes mellitus. METHODS: PCR-RFLP was used to detect the nt3434 A→G variant of mtDNA ND1 gene in 216 diabetic patients and 203 healthy control individuals. Characteristics of mutation and clinical indicators in nt3434A→G family were analyzed. RESULTS: nt3434A→G mutation was detected in one diabetic patient but not found in NC group. This patient had low insulin secretion, low BMI, and elevated serum lactate acids. No significant difference was found in the mutation frequencies between these two groups. nt3434 A→G mutation was also detected in this patient's sister and daughter, who were normal glucose tolerance and had slightly elevated serum lactate acids levels. CONCLUSIONS: Further investigation would be helpful to answer whether nt3434A→G mutation of mitochondrial DNA ND1 gene is associated with an increased risk of diabetes.

[Study on positive rate of blocking antibody in women with recurrent spontaneous abortion administered by route and frequency of paternal lymphocyte immunotherapy].

OBJECTIVE: To investigates factors affecting the positive rate of blocking antibody treated by paternal lymphocyte immunotherapy in patients with recurrent spontaneous abortion (RSA). METHODS: From January 2008 to August 2012, 326 RSA cases undergoing treatment in Infertility Center of Qilu Hospital were studied retrospectively. Those patients were divided into 2 groups randomly: 260 cases in intradermal injection group were administered via bilateral forearm intradermal injections for immunotherapy once 21 days, then the blocking antibody was determined after 2 (23 cases) , 3(73 cases), 4 (74 cases) , 5(90 cases) times respectively, while in subcutaneous injection group, the 66 cases were administered via subcutaneous injection once 21 days, the blocking antibody measured after 3 times; In both cases, the blocking antibody was all determined 2 weeks later. The positive rate of blocking antibodies and the rate of successful pregnancy was recorded, and then followed up after the blocking antibody turning positive. RESULTS: (1) Positive rate of blocking antibodies:the positive rate of blocking antibodies were 17% (4/23) , 58% (42/73), 72% (53/74) and 84% (76/90) in the 2, 3, 4, and 5 times of intradermal injection group, respectively (P < 0.05). In subcutaneous injection group, the positive rate of blocking antibodies was 38 % (25/66), which was significantly lower than that in group intradermal injection receiving 3 times immunotherapy (P < 0.05). (2) The rate of pregnancy:the 176 patients out of 200 patients were pregnant when antibody was positive after immunotherapy, with 71.6% (126/176) of patients gained successful pregnancy(the length of pregnancy more than 5 months). CONCLUSIONS: The route and frequency of administration of immunotherapy could influence the positive rate of blocking antibody. The rate of successful pregnancy will be increased after blocking antibody turning positive.

[The changes of gastrointestinal hormones GLP-1, PYY and ghrelin in patients with newly diagnosed type 2 diabetes mellitus].

OBJECTIVE: To study the changes of plasma glucagon-like peptide-1 (GLP-1), serum peptide-YY (PYY) and Ghrelin and their secretion functions in patients with newly diagnosed type 2 diabetes mellitus (T2DM). METHOD: A total of 102 subjects were enrolled, including 32 normal-glucose-tolerance controls (NGT) and 70 patients with newly diagnosed T2DM. Height, body mass, waist circumference (WC) and hip circumference were measured. The plasma lipids and 0 h, 1/2 h, 2 h plasma glucose, insulin (INS), GLP-1, serum PYY and Ghrelin in a standard meal test in each subject were detected, and body mass index (BMI), homeostasis model assessment of insulin resistance (HOMA-IR), insulin sensitivity index (ISI), homeostasis model assessment of beta cell function (HOMA-B) and early insulin secretion function index (DeltaI30/DeltaG30) were calculated. All these variables were compared between the two groups. RESULTS: Compared with those in NGT group, the WC, fasting plasma glucose (FPG), postprandial plasma glucose (2 h-PG), triglyceride (TG), HOMA-IR were significantly higher (P 0.05), while INS(30), HOMA-B, ISI, DeltaI30/DeltaG30 were significantly lower in T2DM group (P<0. 05). In addition, in T2DM group, 0 h, 1/2 h, 2 h plasma GLP-1 and serum PYY and the area under the curve (AUC) of GLP-1 (GLP-lAuc ) and PYY (PYYAc) in standard meal test were significantly lower (P<0. 05), but the serum Ghrelin and GhrelinA, were significantly higher (P<0. 05). Meanwhile, the secretory peak of GLP-1 and PYY after standard meal in T2DM patients all disappeared. In T2DM group, PYYAUC and TG were negatively correlated (P<0.05), the fasting serum Ghrelin level was negatively associated with total cholesterol (TC), and GhrelinAuc was positively associated with HOMA-B, but negatively with the low-density lipoprotein cholesterol (LDL-C) and FPG (P(<0. 05). CONCLUSION: Patients with newly diagnosed T2DM have decreased fasting and postprandial GLP-1 and PYY levels, along with changes of their secretion mode and increased levels of Ghrelin.

[The efficacy and safety of recombinant human insulin injection in the treatment of diabetic patients:a multicenter, randomized, controlled and open-labeled clinical trial].

OBJECTIVE: A multicenter, randomized, controlled and open-labeled clinical trial was performed to compare the efficacy and safety of recombinant human insulin injection (Yousilin R) and Novolin R in diabetic patients. METHODS: A total of 211 cases were randomized into two groups (1:1) treated with Yousilin R versus Novolin R for 12 weeks respectively. RESULTS: Compared with baseline, the levels of glycosylated hemoglobin A1c (HbA1c) at the end of 12 weeks treatment decreased from 10.77% to 7.72%(P < 0.05) in Yousilin R group and from 10.33% to 7.62% (P < 0.05) in Novolin R group, 2-hour postprandial plasma glucose (2hPG) decreased from 15.49 mmol/L to 9.72 mmol/L (P < 0.05) in Yousilin R group and from 15.33 mmol/L to 10.07 mmol/L (P < 0.05) in Novolin R group, and fasting plasma glucose (FPG) decreased from 10.90 mmol/L to 7.31 mmol/L (P < 0.05) in Yousilin R group and from 10.22 mmol/L to 7.21 mmol/L (P < 0.05) in Novolin R group. The changes of HbA1c, 2hPG and FPG from baseline to endpoint in Yousilin R group was similar to those in Novolin R group (P > 0.05). Furthermore, hypoglycemic events (26.42% vs 30.48%), other adverse events (13.21% vs 16.19%), and serious adverse events (1.89% vs 1.90%) were comparable between Yousilin R and Novolin R groups (P > 0.05). CONCLUSIONS: Yousilin R has similar efficacy, safety and compliance profiles to Novolin R group in the treatment of diabetic patients.

[Molecular scanning of mtDNA gene in two early-onset diabetic pedigrees].

OBJECTIVE: To search for new potential diabetogenic mtDNA defects by scanning mtDNA genome in mitochondrial DNA diabetes (MDM) pedigrees. METHODS: Blood samples were collected from the family members in two suspected MDM pedigrees, which were both maternal transmitted and early-onset diabetes. The whole mtDNA genome except D-loop was detected by direct sequencing in probands of these two diabetic pedigrees. Novel mutations displayed by direct sequencing were then screened in 200 normal glucose tolerant controls and 100 early-onset diabetic patients. RESULTS: We found a novel nt14319T --> C mutation in No. 2 pedigree, but no pathogenic mutation was found in No. 1 pedigree. The mutation of nt14319T --> C, which not being reported previously, locate in the region of ND6 subunit, causing amino acid change (asparagine --> aspartic acid). The frequencies of 14319T/C substitution in 100 early-onset diabetic patients and 200 control subjects were 6% and 5%(P > 0.05). CONCLUSION: A novel mutation 14319T --> C was identified in No. 2 pedigree. All three diabetic family members in this pedigree haboured 14319T --> C mutation, indicating that it may be the major pathogenic mutation for this family.

[Prevalence of diabetes and prediabetes mellitus in the first-degree relatives of patients with type 2 diabetes in Chengdu].

OBJECTIVE: To investigate the prevalence of diabetes and prediabetes mellitus in the first-degree relatives (FDR) of patients with type 2 diabetes (T2DM) in Chengdu. METHODS: A cross-sectional study was undertaken in Chengdu. A total of 2306 adults were recruited, including 535 FDR of T2DM patients and 1771 people without a family history of diabetes. All participants received glucose tolerance tests and measurements of waist, blood pressure and blood lipids. RESULTS: (1) The FDR of T2DM patients had greater standardized prevalence of diabetes than those without a family history of diabetes (26.6% vs. 9.2%). The standardized prevalence of prediabetes in these two groups was 15.0% and 14.1%, respretively. (2) Greater standardized prevalence of diabetes were found in both female (25.5%) and male (28.5%) FDR of T2DM patients compared with their counterparts without a family history of diabetes (women 8.7%, men 11.2%). The standardized prevalence of prediabetes between those with and without a family history of diabetes was 15.9% and 13.4% in women, 13.7% and 15.3% in men, respretively. (3) The younger than 40 years old FDR of T2DM patients had greater prevalence of diabetes and prediabetes than their counterpart without a family history of diabetes, while the FDR of T2DM with an age of > or =40 years old had greater prevalence of diabetes than their counterparts only (P > 0.05). The FDR of T2DM patients with <25 kg/m2 body mass index (BMI) had greater prevalence of diabetes and prediabetes than their counterparts without a family history of diabetes (25.1% vs. 7.4%, 13.2% vs. 9.3%, P < 0.05). The FDR of T2DM patients with > or = 25 kg/m2 BMI had greater prevalence of diabetes (33.0% vs. 13.7%, P < 0.05) but less prevalence of prediabetes (19.2% vs. 26.8%, P < 0.05) than their counterparts without a family history of diabetes. (4) The logistic regression showed that triglyceride (TG) was a risk factor for diabetes in those FDR of T2DM patients (OR = 1.363) and those without a family history of diabetes (OR = 1.27), and high density lipoprotein cholesterol (HDL-C) was a protective factor for diabetes in those without a family history of diabetes (OR = 0.546). CONCLUSION: The FDR of T2DM patients have high risk of diabetes and those younger than 40 years or with <25 kg/m2 BMI also have high risk of prediabetes.

[PPARGC1A gene Thr394Thr G/A polymorphism and its association with type 2 diabetes mellitus and insulin resistance in a Chinese population].

OBJECTIVE: To examine the association of the Thr394Thr polymorphism of PPARGC1A gene with type 2 diabetes (T2DM), insulin resistance (IR) and other metabolic disorders in a Chinese population. METHODS: Three hundred and seven subjects including 151 T2DM patients and 156 normal glucose tolerant controls (NC) were enrolled in this study. The Thr394Thr G/A polymorphism was genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Glucose, insulin, lipids levels were determined in all subjects. Body mass index (BMI), waist circumferences, index of homeostasis model assessment-insulin resistance (HOMA-IR) and blood pressure were also measured. RESULTS: The diabetic subjects had higher levels of BMI, waist circumferences, blood systolic pressure, triglycerides and lower levels of high density lipoprotein-cholesterol (HDL-C) compared with those of control subjects (P<0.05). About 43.7% (66/151) of the T2DM subjects had the AG genotype, while 37.2% (58/156) in the NC group. The frequency of the A allele was 0.225 in T2DM, and 0.186 in the NC subjects. There were no significant differences either in genotype or allelic distribution of G/A polymorphism between the two groups. In the T2DM group, subjects with AA and GA genotypes had significantly higher levels of HOMA-IR, waist circumferences and lower levels of HDL-C (P<0.05) than those carrying GG genotype. HOMA-IR in subjects with AA and AG were significantly higher than those with GG genotype in both groups. CONCLUSION: The A allele of the Thr394Thr (G-->A) polymorphism of the PPARGC1A gene was associated with insulin resistance, and may be related to central obesity and decreased HDL-C levels in Chinese population. The relationship between this polymorphism and T2DM needs further investigation.

[Epidemiological study on metabolic syndrome in Chengdu adult in 2007].

OBJECTIVE: To investigate the prevalence of metabolic syndrome (MS) and associated factors in the adults in Chengdu. METHODS: A cross-sectional survey was undertaken in 2242 residents over 20 years of age. The cluster sampling method was employed to recruit participants from Yulin and Longquan communities in Chengdu. The metabolic syndrome was defined by IDF 2005. RESULTS: The prevalence of MS was 17.2%, with 15.0% and 18.6% for male and female, respectively. The age-adjusted rate of MS was 13.47%, with 17.71% and 11.09% for male and female, respectively. The difference in prevalence of MS between male and female was significant. The prevalence of central obesity, hypertension, hyperglycemia, high triglycerides, low HDL-C were 31.6%, 32.0%, 16.5%, 30.4%, and 31.2% respectively, which became 26.3%, 26.0%, 12.9%, 26.9%, and 30.5% respectively after age-adjustment. Gender differences appeared in all of the components of MS except for low HDL-C (P < 0.05). The multivariable stepwise analysis extracted age, history of hypertension, blood pressure, waist circumference, fasting glycaemia level and triglyceride as major risk factors for MS and blood HDL-C level as a protective factor of MS. CONCLUSION: The prevalence of MS is considerably high in Chengdu. It will continue to rise thanks to the social economic development and population ageing. There is an urgent need to take actions to reduce the burden of MS.

[Glycemic excursions in people with normal glucose tolerance in Chengdu].

OBJECTIVE: To investigate the characteristics of glycemic excursions in people with normal glucose tolerance (NGT) in Chengdu. METHODS: A total of 50 non-obese people with normal glucose tolerance (NGT, 23-68 years old), normal blood pressures and lipid profiles participated in the study. The fluctuations of glucose levels in the participants were measured by a continuous glucose monitoring system (CGMS) for three days 72 h. The 48 h mean blood glucose (MBG), mean amplitude of glycemic excursions (MAGE), Largest amplitude of glycemic excursions (LAGE), Postprandial peak glucose (PPG), Postprandial glucose excursion (PPGE), Mean of postprandial glucose excursion (MPPGE), and absolute means of daily differences (MODD) were measured. RESULTS: The number of glucose values detected by CGMS amounted to 861+/-7 with a mean absolute difference (MAD) of 11.3%+/-10.6%. The CGMS values were significantly correlated with the capillary glucose measurements (n=1076, r=0.761, P<0.005). The participants had a MBG of (5.9+/-1.2) mmol/L, a MAGE of (1.7+/-0.7) mmol/L, a LAGE of (4.4+/-1.9) mmol/L, a daily glycemic peak (PPG) of (8.7+/-1.7) mmol/L, a nadir level of (4.3+/-0.7) mmol/L, a MPPGE of (2.3+/-1.6) mmol/L, and a MODD of (0.75+/-0.79) mmol/L. The post-breakfast Postprandial glycemic excursions (PPGE) were lower than those of post-lunch and post-dinner (P=0.01 and P=0.05). The postprandial glucose excursions in the 60-70 year-old participants were the highest (P<0.022). In 95% (77%-100%) of the daytime, the glycose levels fluctuated between 4.1 and 8.8 mmol/L, and 78% of the participants (n=39) had hyperglycemia (BG>7.8 mmol/L) and 10% (n=5) had asymtomatic hypoglycemia (BG<2.8 mmol/L). CONCLUSION: CGMS tests may be important for detecting asymptomatic hyperglycemia and hypoglycemia. The NGT people in Chengdu have exhibited abnormal blood glucose values in CGMS, revealing problems in people with normal range of blood glucose.

[Response of cryopreserved ovarian tissue after autologous implantation in mouse stimulated with gonadotrophin].

OBJECTIVE: To investigate the response of cryopreserved ovarian tissue after autologous implantation in mouse stimulated with gonadotrophin. METHODS: Thirty six female mice were randomly divided into three groups, with 12 mice in each group. In group of fresh ovarian tissue, fresh ovarian tissue was implanted into kidney capsule of mice; in group of cryopreserved ovarian tissue, ovarian tissue was implanted into kidney capsule of mice after cryopreserved by vitrification for two weeks. We investigated the response of ovarian tissue two weeks later after autologous implantation stimulated with gonadotrophin. Immunohistochemistry staining method was used to observe the expression of follicle stimulating hormone receptor. RESULTS: Before and after stimulation with gonadotrophin, the mature follicle rate of group of fresh ovarian tissue was 2.3% and 4.2%, that of group of cryopreserved ovarian tissue was 2.3% and 4.0%, and that of group of control was 2.6% and 5.8%. Regarding the percentages of mature follicle, there were significant differences after stimulation with gonadotrophin (P < 0.05). After stimulating with gonadotrophin the percentages of mature follicle were the same in the fresh tissue group, cryopreserved tissue group and control group (P > 0.05). The integrated optical density of follicle stimulating hormone receptor of fresh ovarian tissue in antrofollicle and pre-antrofollicle were 9408 +/- 2777 and 4531 +/- 1903, that of cryopreserved ovarian tissue were 9175 +/- 3093 and 4808 +/- 1386, and that of the control ovarian tissue were 8838 +/- 2064 and 5516 +/- 1136 respectively. There was no significant difference between any two groups (P > 0.05). CONCLUSION: The follicle stimulating hormone receptor is preserved by cryopreservation and transplantation, small pieces of ovarian tissue response to gonadotropin stimulation is normal.

[Capability of oocyte maturation in human cryopreserved ovarian tissue following xenografting].

OBJECTIVE: To investigate the development and maturation competence of oocytes retrieved from cryopreserved and transplanted human fetal ovarian tissue by techniques of tissue culture, inducing ovary, oocyte retrieval, and in vitro maturation (IVM). METHODS: Fetal ovaries of 20 weeks were frozen-thawed and cultured for 6 days in vitro, then xenografted into kidney capsules of immunodeficient mice. All mice were stimulated with follicle stimulating hormone every second day for 23 weeks, starting 1 week after grafting. Then oocytes were retrieved from antral follicles 13 hours after human chorionic gonadotrophin injection. IVM was performed to evaluate the maturation competence of the oocytes from ovarian grafts. Human fetal ovarian tissues were examined with histological and proliferating cell nuclear antigen (PCNA) evaluation. RESULTS: There was no difference between fresh ovarian tissues and frozen-thawed ovarian tissues in the percentage of follicles at different growth stages (P > 0.05). The proportion of the primary follicles and preantral follicles in the cultured ovarian tissues was significantly larger than that of fresh ovarian tissues and frozen-thawed ovarian tissues (P < 0.05). The proportion of the primary follicles, preantral follicles, and antral follicles in the transplanted ovarian tissues was significantly higher than that of cultured ovarian tissues, fresh ovarian tissues, and frozen-thawed ovarian tissues (P < 0.05). No significant signals of PCNA in the primordial follicles in all ovarian tissues were observed. PCNA immunoreactivity first appeared in primary follicles. However, the obviously positive signals of PCNA were seen in the oocytes and/or the granular cells of cultured ovarian tissues and transplanted ovarian tissues. Oocytes from antral follicles were collected and matured in vitro, and 21.43% of the oocytes reached to MII within 48 hours IVM. CONCLUSIONS: Human ovarian follicles can survive and develop well after cryopreservation, tissue culture, and xenotransplantation. Furthermore, oocytes recovered from grafts have normal maturation competence.

[Relationship between peripheral insulin resistance and beta-cell function in obese subjects].

OBJECTIVE: To detect the relationship between peripheral insulin resistance and beta-cell function in obese subjects with normal blood glucose, and to explore the role of free fatty acids (FFAs) in the pathogenesis of type 2 DM. METHODS: Ten simple obesity male subjects (OB) with normal blood glucose and 10 lean male persons (NC) matched on age and living style were included. None of them was smoker, and their blood pressure, serum cholesterol levels were all normal. All subjects underwent the following tests: (1) Fasting serum lipids profile and FFA; (2) Baseline immunoreactive insulin (IRI), pro-insulin (PI) and true-insulin (TI); (3) Acute insulin response (AIR) to an intravenous bonus of glucose; (4) Euglycemic hyperinsulinemia clamp study to assess the peripheral glucose disposal rate (GDR). RESULTS: In obesity subjects, the baseline IRI, PI and TI were elevated and the ratio of PI/(PI + TI) was higher than controls (P < 0.01). The AIR was enhanced in OB and the area under the curve was larger (P < 0.05), but in obesity subjects the plasma glucose levels at 3, 5 and 10 minutes were much higher than controls (P < 0.05). The GDRs obtained during steady state were markedly decreased in OB (P < 0.05). The serum lipids profiles were normal except that triglyceride was higher in OB (P < 0.05) and the FFAs concentration was also elevated in OB (P < 0.05). There was a negative correlation between FFAs concentration and GDR in OB groups (r = -0.767, P < 0.01). CONCLUSION: The present data showed that in obese subjects with normal blood glucose, there were severe peripheral insulin resistance, relative deficiency of beta-cell first-phase insulin release and increased ratio of pro-insulin to insulin. It is logical to deduce that both insulin resistance and the beta-cell changes could be, at least partially, secondary to high levels of serum FFAs. Elevated circulating FFAs may paly a role in the pathogenesis of type 2 DM.

[The relationship between insulin resistance and endothelium-dependent vasodilatation in obese subjects].

OBJECTIVE: To study the relationship between insulin resistance and endothelium-dependent vasodilatation function in obese subjects with normal blood glucose, serum cholesterol and blood pressure and to explore the non-traditional risk factors associated with atherosclerosis. METHODS: 24 euglycemic obese males (OB) and 12 age and sex matched lean controls (Lean) underwent oral glucose tolerance test (OGTT), euglycemic hyperinsulinemia clamp study with an insulin infusion rate of 120 mU/m(2)/min to evaluate the peripheral glucose disposal rate (GDR) in steady-state, brachial artery ultrasound study to assess the endothelium-dependent vasodilation (EDV) and independent vasodilation (EIV), the combined intima-media thickness (IMT) of common carotid arteries examined by high resolution ultrasound, and fasting lipids profile and free fatty acids (FFAs) concentration. RESULTS: The blood glucose and serum cholesterol levels were all normal in the OB group but the serum FFA and triglyceride concentrations were elevated compared with those of the controls (FFA: 888 microM/l +/- 158 microM/l vs 508 microM/l +/- 137 microM/l, P < 0.05; TG: 1.8 mM/l +/- 0.6 mM/l vs 1.1 mM/l +/- 0.5 mM/l, P < 0.05) The GDR in steady-state was 6.7 mg x min(-1) x kg(-1) +/- 1.4 mg x min(-1) x kg(-1) in the OB group, significantly lower than that of the controls (12.2 mg x min(-1) x kg(-1) +/- 3.1 mg x min(-1) x kg(-1), P < 0.05). In the OB group the flow-mediated dilation of the brachial artery caused by reactive hyperemia (EDV) were impaired and the relaxation time was shorter than that of the controls (P < 0.05) but the EIV was not significantly different between these two groups. There was a significantly positive correlation between GDR and EDV (r = 0.438, P < 0.05). The IMT of common carotid arteries was thicker in the OB group than in the control group (0.50 mm +/- 0.02 mm vs 0.34 mm +/- 0.19 mm). CONCLUSION: The endothelium-dependent vasodilation was impaired in obese subjects even though without hyperglycemia, hypercholesterolemia, and hypertension. Traditional cardiovascular risk factors cannot fully account for the increased risk of cardiovascular disease in metabolic syndrome. Elevated circulating FFA concentration, is characteristic of metabolic syndrome, may play a role in endothelial dysfunction.

Short-term glucose metabolism and gut hormone modulations after Billroth II gastrojejunostomy in nonobese gastric cancer patients with type 2 diabetes mellitus, impaired glucose tolerance and normal glucose tolerance.

BACKGROUND AND AIMS: Roux-en-Y gastric bypass (RYGB) is effective in controlling blood glucose in obese patients with type 2 diabetes (T2DM). The alterations of gut hormones involving in glucose metabolism may play an important role. Our aim was to explore the short-term effects of Billroth II gastrojejunostomy (a similar type of RYGB) on glucose metabolism and gut hormone modulations in nonobese patients with different levels of blood glucose tolerance. METHODS: Twenty one nonobese gastric cancer patients with different levels of blood glucose tolerance were submitted to Billroth II gastrojejunostomy. Among them, seven had T2DM, seven with impaired glucose tolerance (IGT) and the other seven had normal glucose tolerance (NGT). Body weight, glucose parameters, responses of plasma glucagon-like peptide-1 (GLP-1), peptide YY (PYY) and gastric inhibitory polypeptide (GIP) to 75 g glucose were measured at baseline and 3 months after surgery. RESULTS: Similar weight losses were observed in all groups. Blood glucose was reduced in T2DM and IGT patients. Fasting and 30-min plasma glucose were increased significantly in NGT. GLP-1 showed insignificant alterations in all groups. PYY was evaluated in T2DM and IGT but remained unchanged in the NGT group. Decreased fasting and AUC GIP were observed in patients with T2DM; however, fasting and 30-min GIP were increased in NGT patients. CONCLUSIONS: Billroth II gastrojejunostomy is effective in reducing blood glucose in nonobese patients with T2DM and IGT but could deteriorate early blood glucose in nonobese NGT in a 3-month time period. Variations of glucose and gut hormone changes in the three groups suggest a role of proximal intestine in the pathophysiology of T2DM.

[Therapeutic effects of bone marrow transplantation on ovarian injury in mice].

OBJECTIVE: To evaluate the therapeutic effects of bone marrow transplantation (BMT) on ovarian injury induced by chemotherapy in mice. METHODS: Forty-eight mice were randomized equally into normal control group (A), cyclophosphamide and BMT group (B), and cyclophosphamide group (C). The mice in groups B and C were treated with intraperitoneal injection of cyclophosphamide at the daily dose of 150 mg/kg for 3 consecutive days, and allogeneic bone marrow cell transplantation was performed in group B. The ovary coefficient and the amount of follicles were compared to evaluate the function of ovaries. For cell tracking, the bone marrow cells were labeled with Hoechst 33342 and detected through fluorescence microscope after transplantation. RESULTS: On days 21 and 50 after cyclophosphamide treatment, the ovary coefficient and the amount of follicles were significantly lowered in groups B and C (P<0.05), but the reduction was obviously ameliorated in group B (P<0.05). Cell tracking showed the presence of the donor bone marrow cells in the ovaries of the recipients mice after BMT. CONCLUSION: BMT can improve the ovarian function impaired by chemotherapy in mice.

Effect of different sites for cryopreserved ovarian tissue implantation in rabbit.

BACKGROUND: Autotransplantation of frozen-thawed ovarian tissue has proven to be an effective method to restore endocrine function and fertility. But it remains to be studied which site and which method is most effective and practical. We therefore implanted small pieces of cryopreserved ovarian tissues into different sites in rabbits to find the optimal position. METHODS: Fifteen New Zealand white female rabbits were randomly divided into three groups. In group 1, fresh ovarian tissues were implanted into the mesometrium and ovarian bursa. In group 2, cryopreserved ovarian tissues were implanted into the mesometrium and ovarian bursa. In group 3, cryopreserved ovarian tissues were implanted into the preserved ovary. RESULTS: There were no significant differences among the three groups as to the proportions of normal and morphologically changed follicles in implanted ovarian tissues. The implanted ovarian tissues in the three groups did not show any evident changes in histology and ultrastructure, and all resumed follicle development and revealed maturescent follicles. CONCLUSIONS: Cryopreservation and implantation of small pieces of ovarian tissues are feasible. Generally, the mesometrium, ovarian bursa and ovary are all available sites for implantation and have similar rates of acceptance, despite some differences in the details of implantation.