Objective To observe the effect of metformin and exercise therapy for IGT. Methods 86 patients with IGT were given metformin and exercise therapy,the range of glucose,BMI and symptoms improvement were observed before and after the therapy. Results After 6 months therapy,the fasting blood glucose and 2 h after oral administration of 75 g glucose levels were significantly reduced(P＜0.01＝,BMI reduced(P＜0.05＝.72 cases (83.7%)were back to normal glucose tolerance,14 cases maintained IGT(those unable to adhere to exercise and no diet controlled),0 case became DM. Conclusion Metformin and exercise therapy had good efficacy in curing IGT.

Objective To clone the human Islet neogenesis associated protein(rhINGAP)gene,express the gene extraeellulary in Pichia yeast for.further study on biological function and animal test on INGAP.Methods INGAP gene Was amplified with PCR and inserted into the recombinant plasmidα/pUC18.Then,the fusion gene of α and INGAP was digested and inserted into the expression plasmid pPIC9K.The positive recombinant plasmid which integrated INGAP Was confirmed by restriction enzyme digestion and sequencing,and it Was linearized with Sal Ⅰ digestion and transfered into the yeast host strain GS115 through electroporation.The yeast transformants that harbor the desired gene INGAP with high copy were selected by the auxotroph mediam G418,and verified by PCR.The condition of hake-flask culture was optimized,and the recombinant human INGAP Was induced expression with methanol as the only Carbone source.The antigen activity of the desired protein Was detected by Western blotting and ELISA method.Results Recombinant plasmid αINGAP/pPIC9K were successfully constructed and three positive Pichia yeast transformants were obtained.The expressed protein had satisfactory antigen activity,which Was confirmed by the Western blotting and ELISA method.Conclusions Pichia yeast expressing human Islet neogenesis associated protein (rhINGAP)gene was successfully constructed.

Objective To investigate the effects on self-renewal of pancreatic cancer stem cells by inhibiting hedgehog signaling pathway through cyclopamine. Methods PANC1 stem cells, PANC1 adherent cells and immortalized pancreatic ductal epithelial H6C7 cells were treated with 0.5, 1, 2, 5, 10 mol/L of cyclopamine for 24, 48, 72 h. The expression of Smo mRNA and Gli1 mRNA were detected by real-time PCR.Cell growth viability was measured by CCK 8. Cell cycle and apoptosis were determined by flow cytometry.Results Seventy-two hours after cyclopamine treatment, the Smo mRNA expressions of PANC1 stem cells,PANC1 adherent cells and H6C7 cells were 1,0.83 and 2.61; the expressions of Gli mRNA were 57.27,26.35,1; the inhibitory rates were ( 37.85 ± 13.69 ) %, ( 8.53 ± 4.43 ) %, (43.55 ± 28.98 ) %. Compared with PANC1, the expressions of Smo mRNA, Gli1 mRNA and the inhibitory rate of PANC1 stem cells significantly increased ( P ＜ 0.05 ). The proportion of G1 stage of PANC1 stem cells significantly decreased from (67.41 ±6.35)% to (36.53 ±6.03)% (P ＜0.05), and the apoptosis decreased from (10.95 ±5.68) % to ( 5.73 ± 1.42 ) % ( P ＞ 0.05 ). The proportion of G1 stage of PANC1 cells significantly decreased from ( 67.64 ± 6.88 ) % to ( 53.13 ± 1.10 ) % ( P ＜ 0.05 ); the apoptosis decreased from ( 12.08 ±4.12)% to (5.66 ± 1.33)% (P ＞0.05). While both the proportion of G1 stage and apoptosis of H6C7 cells was not significantly different. Conclusions Cyclopamine can inhibit the proliferation of PANC1 stem cells via blocking hedgehog signal pathway, and the mechanism may not be associated with cell apoptosis.

Objective To establish the method of suspension culture for stem cells from human pancreatic cancer cell line PANC1.Methods PANC1 cells were cultured in serum-free medium under floating-culture system.Tumor cell spheres were observed by optical microscope.Expression of CD133 and cell cycle were detected by flow cytometry.Cancer stem cells were induced to differentiate with 10%FBS,and expression of CK18,was evaluated with immunofluorescence microscope.Spheres cells were injected into the subcutaneous space of NOD/SCID rat and tumor formation was monitored weekly.Results PANC1 cells could form the stem cells spheres,and the rate of sphere formation was stable between 4%0 and 5%0 after 20 passages in vitro.The expression of CD133(5.91±0.7%)and proportion of G0/G1 phase cell(80.99±2.60%)was significantly increased in spheres cells compared with parental PANC1 cells(1.44±0.52%and 69.01±5.03%),and the difference was statistically significant(P＜0.05).When these spheres cells were cultured in media with serum,these cells gradually returned to the status of parental cells and expressed CK18,2×103 sphere cells injection could initiate tumor fornmtion in NOD/SCID rat.Conclusions Tumor spheres stem cellscould be generated under serum-free floating-culture system.The sphere cells possessed the capacities of self renew,difierentiation,and tumorigenic potential.

Objective To investigate the incidence of newly-onset type 1 diabetes mellitus ( T1DM ) complicated with ketoacidosis(DKA) and its relevant factors in pediatrics. Methods Hospital records of 317 T1DM children below 18 years of age, diagnosed from 2010 to 2012 were reviewed. By using retrospectively analyzed data of inpatients with newly-diagnosed T1DM, the incidence of DKA was calculated. In this study, the influential factors of DKA included gender, age, residence, family history of diabetes mellitus, duration of symptoms, misdiagnosis or missed diagnosis, and delayed treatment. Patients were divided into two groups:group 1, aged<5 year and group 2, aged>5 year. Results Of all patients diagnosed with T1DM, 175 ( 55. 2%) presented with DKA, and mild, moderate, and severe DKA accounted for 26. 5%, 23. 9%, 49. 6%, respectively. The incidences of DKA in group 1 andgroup2were67.5% and48.0% (P=0.001),withthehighestfrequency(70.3%)inpatientsaged<2 years. The proportion of severe DKA in group 1 was significantly higher than that of group 2 (60. 0% vs 41. 3%, P=0. 048). The rates of misdiagnosis and missed diagnosis in the two groups were respectively 27. 4% and 12. 0%(P=0. 001), being 37. 8% in children<2 years. The HbA1C level of group 1 was lower than group 2 (11. 50% vs 12.54%,P=0.001). Intheacutemetabolicandhoneymoonperiod,Cpeptidelevelsofgroup1werebothlowerthan those of group 2 [(0. 36 vs 0. 55) ng/ml, P=0. 001;(0. 40 vs 0. 61) ng/ml, P=0. 02]. The DKA incidence of patients with misdiagnosis or missed diagnosis was significantly increased(83. 9% vs 49. 0%, P=0. 000). Compared with those without DKA, C peptide level of patients with DKA was lower in the acute metabolic period[(0. 56 vs 0.40)ng/ml,P<0. 01], but no difference in honeymoon period[(0. 67 vs 0. 59)ng/ml,P=0. 22]. Logistic regression showed that age, misdiagnosis or missed diagnosis were associated with the presence of DKA. The possibility of the occurrence of DKA in patients aged>5 years was half of patients aged<5 years ( OR=0. 448, P=0. 003), and the risk of DKA in patients with misdiagnosis or missed diagnosis was higher (OR=5. 640, P=0. 005). Conclusion DKA in patients with newly-onset T1DM is frequent and often severe. Multivariate analysis revealed that patients aged <5 years and those with misdiagnosis or missed diagnosis are encountered high risk of DKA.

Objective:To investigate the prognostic significance of D-dimer level in patients with diffuse large B-cell lymphoma (DLBCL).Methods:The clinical data of 70 newly diagnosed DLBCL patients who were admitted to Tianjin People's Hospital from January 2015 to June 2019 were retrospectively analyzed. The optimal cut-off value of D-dimer for survival was determined according to the receiver operating characteristic (ROC) curve, and the patients were grouped. The differences of coagulation related indexes and clinicopathological features between patients with different D-dimer levels were compared. Kaplan-Meier method was used for univariate analysis of overall survival (OS), and Cox regression model was used for multivariate analysis of OS.Results:According to ROC curve, the best cut-off value of D-dimer for survival was 0.75 mg/L. The proportion of patients with different clinical staging, international prognostic index score, lactate dehydrogenase level had statistically significant differences between the D-dimer ≥0.75 mg/L group (36 cases) and <0.75 mg/L group (34 cases) (all P < 0.05). The prothrombin time of D-dimer ≥ 0.75 mg/L group and < 0.75 mg/L group were (13.5±0.9) s and (13.0±0.8) s, respectively, and the activated partial thromboplastin time were (37±5) s and (34±6) s, respectively,and the differences were statistically significant (all P < 0.05). Univariate analysis showed that the 5-year OS rates of DLBCL patients with Ann Arbor stage Ⅲ-Ⅳ, international prognostic index score > 2, lactate dehydrogenase level > 240 U/L, B symptoms, D-dimer level ≥0.75 mg/L were decreased (all P < 0.05). Multivariate Cox regression analysis showed that D-dimer ≥0.75 mg/L was an independent risk factor for OS of DLBCL patients ( HR=0.368, 95% CI 0.144-0.944, P= 0.038). Conclusion:The level of D-dimer can be used as a clinical indicator to judge the prognosis of DLBCL patients, and the prognosis of patients with high D-dimer level is poor.

Objective To investigate the expression of high?molecular?weight keratins CK5/6, CK14, estrogen receptor ( ER) and progesterone receptor ( PR) in differential diagnosis of simple ductal hyperplasia ( UDH) , atypical ductal hyperplasia ( ADH) and low?grade ductal carcinoma in situ ( low?grade DCIS) . Methods The clinicopathological data of twenty cases of atypical ductal epithelial hyperplasia ( ADH) with focal cancerization changed into low?grade DCIS diagnosed at Tianjin Medical University Cancer Institute and Hospital between January 2013 and February 2014 were reviewed and analyzed. The expressions of CK5/6, CK14, ER and PR were detected by immunohistochemistry. Results Positive expressions of CK5/6 and CK14 were seen in UDH showing a mosaic pattern, while negative expression in ADH and low?grade DCIS. In addition, CK5/6 and CK14 were positively expressed in the myoepithelial cells of UDH, ADH and low?grade DCIS. Positive expressions of ER and PR were observed in UDH, ADH and low?grade DCIS. But they presented diffuse and homogeneous strong positive expression in ADH and variable positive expression in UDH. Conclusion In the intraductal proliferative lesions of the breast, the use of combined detection of the expression of CK5/6, CK14, ER and PR is of practical significance in the differential diagnosis of UDH, ADH and low?grade DCIS.

Objective To investigate the expression of high?molecular?weight keratins CK5/6, CK14, estrogen receptor ( ER) and progesterone receptor ( PR) in differential diagnosis of simple ductal hyperplasia ( UDH) , atypical ductal hyperplasia ( ADH) and low?grade ductal carcinoma in situ ( low?grade DCIS) . Methods The clinicopathological data of twenty cases of atypical ductal epithelial hyperplasia ( ADH) with focal cancerization changed into low?grade DCIS diagnosed at Tianjin Medical University Cancer Institute and Hospital between January 2013 and February 2014 were reviewed and analyzed. The expressions of CK5/6, CK14, ER and PR were detected by immunohistochemistry. Results Positive expressions of CK5/6 and CK14 were seen in UDH showing a mosaic pattern, while negative expression in ADH and low?grade DCIS. In addition, CK5/6 and CK14 were positively expressed in the myoepithelial cells of UDH, ADH and low?grade DCIS. Positive expressions of ER and PR were observed in UDH, ADH and low?grade DCIS. But they presented diffuse and homogeneous strong positive expression in ADH and variable positive expression in UDH. Conclusion In the intraductal proliferative lesions of the breast, the use of combined detection of the expression of CK5/6, CK14, ER and PR is of practical significance in the differential diagnosis of UDH, ADH and low?grade DCIS.

OBJECTIVETo investigate the expression of high-molecular-weight keratins CK5/6, CK14, estrogen receptor (ER) and progesterone receptor (PR) in differential diagnosis of simple ductal hyperplasia (UDH), atypical ductal hyperplasia (ADH) and low-grade ductal carcinoma in situ (low-grade DCIS) .

METHODSThe clinicopathological data of twenty cases of atypical ductal epithelial hyperplasia (ADH) with focal cancerization changed into low-grade DCIS diagnosed at Tianjin Medical University Cancer Institute and Hospital between January 2013 and February 2014 were reviewed and analyzed. The expressions of CK5/6, CK14, ER and PR were detected by immunohistochemistry.

RESULTSPositive expressions of CK5/6 and CK14 were seen in UDH showing a mosaic pattern, while negative expression in ADH and low-grade DCIS. In addition, CK5/6 and CK14 were positively expressed in the myoepithelial cells of UDH, ADH and low-grade DCIS. Positive expressions of ER and PR were observed in UDH, ADH and low-grade DCIS. But they presented diffuse and homogeneous strong positive expression in ADH and variable positive expression in UDH.

CONCLUSIONIn the intraductal proliferative lesions of the breast, the use of combined detection of the expression of CK5/6, CK14, ER and PR is of practical significance in the differential diagnosis of UDH, ADH and low-grade DCIS.

Breast ; metabolism ; pathology ; Breast Neoplasms ; diagnosis ; metabolism ; Carcinoma, Ductal, Breast ; diagnosis ; metabolism ; Carcinoma, Intraductal, Noninfiltrating ; diagnosis ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Hyperplasia ; diagnosis ; metabolism ; Immunohistochemistry ; Keratin-14 ; metabolism ; Keratin-5 ; metabolism ; Keratin-6 ; metabolism ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism

BACKGROUNDIt has been indicated that autologous hematopoietic stem cell transplantation (AHST) is a promising treatment to adults with type 1 diabetes, however, the application of AHST therapy to children with type 1 diabetes still needs more data. The aim of this study was to assess the clinical effect of immune intervention combined with AHST and conventional insulin therapy in the treatment of children with newly diagnosed type 1 diabetes.

METHODSThis 1:2 matched case-control study was comprised of 42 children who were newly diagnosed with type 1 diabetes in the Department of Endocrinology, Beijing Children's Hospital from 2009-2010. The case group included 14 patients, who were treated with AHST within the first 3 months after being diagnosed with diabetes at request of their parents during 2009-2010. The control group included 28 patients with newly diagnosed type 1 diabetes at the same period of hospitalization. We compared the baseline and follow-up data of them, including ketoacidosis onset, clinical variables (glycosylated hemoglobin (HbA1c), insulin dosage and serum C-peptide).

RESULTSThe clinical characteristics of the patients was comparable between the case group and the control group. At 6-12 months ((10.7±4.2) months) after AHST treatment, we found 11 patients in the case group did not stop the insulin therapy, three cases stopped insulin treatment for 2, 3 and 11 months, respectively. No diabetic ketoacidosis (DKA) occurred after transplantation in all the patients in the case group. HbA1c in the control group was significant lower than that in the case group (P < 0.01), while the insulin dosage and serum C-peptide were not significant different between the two groups (P > 0.05). In order to eliminate the honeymoon effect, we performed final follow-up at the 3-5 years ((4.2±1.8) years) after AHST treatment, and found that HbA1c in the control group was still lower than that in the case group (P < 0.01); however, the insulin dosage and serum C-peptide were not significantly different between the two groups (P > 0.05). Moreover, the insulin dosage was not significant different from baseline to follow-up period in the case group.

CONCLUSIONAHST treatment showed no advantage in effectiveness in children with newly diagnosed type 1 diabetes, both in insulin dose and long term blood glucose control.

Adolescent ; Case-Control Studies ; Child ; Child, Preschool ; Diabetes Mellitus, Type 1 ; drug therapy ; therapy ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Hypoglycemic Agents ; therapeutic use ; Infant ; Insulin ; therapeutic use ; Male ; Transplantation, Autologous ; Treatment Outcome