OBJECTIVE:To investigate the function change of intestinal CYP3A4 and P-glycoprotein (P-gp) under diabetic conditions and its mechanism. METHODS:Caco-2 cells were respectively added with 25, 50 and 100 μmol/L midazolam (CYP3A4 probe substrate) for incubation for 15,30,60,120,180 min;with 0.1,0.2,0.4 μg/ml rhodamine 123 (P-gp sub-strate)for incubation for 15,30,60,90,120 min to determine the substrate concentrations and incubation time. Caco-2 cells were added with insulin,glucose and fatty acid (palmitic acid and oleic acid) at different concentrations,and then the production of 1′-OH-midazolam,the metabolite of midazolam,and the intake of rhodamine 123 were determined,in order to investigated the ef-fect on the function of CYP3A4 and P-gp. RESULTS:The optimal substrate concentrations and incubation time were as follows as 50 μmol/L midazolam,0.1 μg/ml rhodamine 123 and incubation for 2 h. With the increase in the concentration of insulin,glucose and palmitic acid,the production of 1′-OH-midazolam and the activity of CYP3A4 reduced;the intake of rhodamine 123 in-creased,and the efflux transport function of P-gp decreased. Oleic acid had no significant effect on the production of 1′-OH-mid-azolam or the intake of rhodamine 123. CONCLUSIONS:Under diabetes condition,the increase of insulin,glucose and palmitic acid may reduce the function of CYP3A4 and P-gp,while oleic acid has no effect on the function.

Diabetes,characterized by chronic increase of blood glucose,is a group of metabolic diseases.It is usually treated by oral administration with antidiabetic drug and insulin with western medicine.Diabetes is called consumptive thirst in TCM.Professor NAN Zheng thought the pathogenesis of consumptive was yin defi ciency and dryness-heat at the early stage,and at the protracted stage it was usually characterized by qi defi ciency,blood stasis,pathogen intruding into collateral and poison damaging zang and fu.He established a series of treatment plan,involving diet,movement,and the Chinese materia medica.Professor NAN opposed to abusing insulin and the insulin multi-purpose theory,and proposed the treatment should start from change of bad life style and combine with application of Chinese medicine to regulate yin-yang and qi-blood of fi ve viscera,treat triple diabetes at the same time.So long as syndrome differentiation was accurate,the Chinese materia medica may replace the insulin and prolong the life.In conclusion,this article enumerates 3 typical cases diabetes withdrawal of insulin with TCM by Professor NAN Zheng.

To study a method for determination of sulfamethoxazole, sulfadiazine and trimethoprim tablets, a compound tablet of sulfamethoxazole, sulfadiazine and trimethoprim. METHOD: A capillary zone electrophoresis (CZE) method was used to assay three components of this compound preparation. RESULTS: The complete separation of components was achieved with 0.05 molL-1 pH 6.0 running phosphate buffer and a constant voltage of 20 kV (current of 95 μA～105 μA). The retention times of individual components were between three and eight minutes and a good linearity was shown between concentration and peak area in the concentration range over 70 μgml-1～750 μgml-1. When acetanilide was used as internal standard, the relative standard deviation (RSD) of each component determined in a batch was less than 1% (n=9). The recovery of each component was not less than 96.45% with RSD less than 3%. The analytical results obtained from 6 samples of clinical use were inconsistent with those by standard method, however the quantity of each sulfa drug was obtained by CZE method. CONCLUSION: The results showed this method is accurate, simple, and rapid. When this method is used, the quantity of each three components is determined, but by the standard method, only the total quantity of the two sulfa drugs is obtained.

0. 20) after D test of normality. The normal range of GEMP was 63. 11-72. 94 ?mol fructose/L (P= 95%). When clinical cardiovasculopathy, nephropathy, retinopathy,peripheral neuropathy and auditus reduction in the aged were compared between two groups of diabetic patients, one with normal and another with abnormal values of GEMP, the clinical indices, excepting cardiovasculopathy, were significantly different between the groups.

ObjectiveTo established a cell line that expresses hBD1 stably,and detected the antimicrobial activity of the hBD1 to the muhidrug resistant bacterial strains.MethodsRecombinant plasmid was introduced into COS-7 cells by lipofectamine,cells were selected in culture medium containing G418 to acquired the monoclonal cell lines,total RNA were extracted from the cultured cells,expression levels of hBD1 mRNA was identified by RT-PCR,collected the supernatant solution of the cultured cell,expression levels of protein was identified by Western blot.Put the expression products and resistant organisms mixed together,after incubation in different times in 37℃,coating the mixtures in LB flat,then obtained the ratios between colonies number of experimental groups and colonies number of control groups,put those ratios as the survival rate of the drug resistance bacterias.Results The monoclonal cell lines had obtained after screened with G418,the hBD1 gene could be detected both at transcriptional and protein levels,Under the influence of expression product hBD1,survival rate of muhidrug-resistant Acinetobacter baumannii,multidrug-resistant Escherichia coli and multidrug-resistant Klebsiella pneumoniae could reduced to 9%,22% and 50%,but survival rate of multidrug-resistant Stenotrophomonas maltophilia is not have apparente difference with the control group.ConclusionThe stably-transfected cell line of hBD1 was successfully constructed,and the expression products of hBD1 showed the antimicrobial activity toward multidrug resistant bacterial strains.

AIM:To investigate whether the protective effect of adiponectin on glucose and lipid metabolism is achieved through down-regulating major histocompatibility complex classⅡ( MHCⅡ) in the adipose tissue.METHODS:Adiponectin knockout ( KO) mice and C57BL/6 ( WT) mice were fed with high-fat diet and standard diet for 24 weeks, re-spectively.The body weight, fasting blood glucose (FBG), fasting insulin (FINS), homeostasis model assessment of insu-lin resistance (HOMA-IR), triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol ( HDL-C) , hepatic histology, and classⅡtrans-activator ( CIITA) , histocompatibility 2 class II antigen E beta (H2-Eb1) and cluster of differentiation 74 (CD74) mRNA and MHC II protein levels in adipose tissue were measured at sacrifice.siRNA targeting MHC II and overexpression vector was used in 3T3-L1 cells to explore the effect of adiponectin on the protein level of MHCⅡ.RESULTS:The levels of body weight, FBG, FINS, HOMA-IR, TC, TG, LDL-C, hepatic steatosis, CIITA, H2-Eb1 and CD74 mRNA expression, and MHCⅡ protein expression in the KO mice were higher than those in the WT mice that fed with high-fat diet or standard diet.In 3T3-L1 cells, inhibition of adiponectin reversed MHC II protein level induced by specific siRNA.The expression of MHC II in adipocytes decreased after adiponectin was overexpressed.CONCLUSION: Adiponectin improves glucose and lipid metabolism through sup-pressing the expression of MHCⅡin the adipose tissue.

BACKGROUND:The functions of homo heterogeneous ribonucleoprotein E1 are very wide. It can participate in the expression of skeleton proteins in the nervous system. OBJECTIVE:To construct the recombinant plasmid of homo heterogeneous ribonucleoprotein E1 and observe its expression in nerve cells for further studying the functions of it in neurocytes. METHODS:Using pcDNATM4/His C, the homo heterogeneous ribonucleoprotein E1 was subcloned into recombinant plasmid E1-pcDNATM 4/His C, fol owed by enzyming and sequencing. After SH-SY5Y cells were transfected with the recombinant plasmid, western blot analysis and real time RT-PCR were used to detect the expression of homo heterogeneous ribonucleoprotein E1 in SH-SY5Y cells. And the growth of SH-SY5Y cells was observed. RESULTS AND CONCLUSION:We successful y constructed the eukaryotic expressed vector of homo heterogeneous ribonucleoprotein E1. The recombinant plasmids were verified to express in SH-SY5Y cells correctly at mRNA and protein levels. And SH-SY5Y cells generated quickly after homo heterogeneous ribonucleoprotein E1 was over-expressed. The homo heterogeneous ribonucleoprotein E1 is an important protein in neural development. And this vector offers the premise for further studying its function in nervous system.

Background and purpose:P38 mitogen-activated protein kinase (P38MAPK) signal transduction pathway is involved in occurrence, development and transfer process in a wide variety of tumors. Urokinase-type plasminogen activator (uPA) plays an important role in tumor invasion and metastasis. This study aimed to explore the clinical signiifcance of the P38MAPK signaling pathway and the expression of uPA in ovarian cancer.Methods:The expressions of uPA, P38MAPK, ERK and AKT were detected in 49 cases of cervical adenocarcinoma by immunohistochemistry. The expressions of uPA and P38MAPK were detected by Western blot in ovarian cancer cell lines HO8910, HO-8910PM, SKOV3 and CAOV3. The changes of uPA and P38MAPK were detected by SB203580, a speciifc inhibitor of P38MAPK signal transduction pathway.Results:The result of immunohistochemical method showed that positive expression rates for uPA, P38MAPK, ERK and AKT were 61.22%, 57.14%, 53.06% and 55.10%, respectively. The expression of the uPA was positively correlated with the P38MAPK (r=0.865,P=0.001), and related with clinicopathologic stage, differentiated degree, lymph node metastasis, but not related with age and histologic type (P>0.05). The expressions of AKT and ERK were related with the lymph node metastasis and greater omentum metastasis(P<0.05), but not related with age and histologic type (P>0.05). The expression of uPA in HO-8910PM was higher than that in ovarian cancer cell lines HO8910, SKOV3 and CAOV3, and the expression of uPA reduced when the P38MAPK signal transduction pathway was cut off by the SB203580. The expressions of P38MAPK and uPA were negatively correlated with the prognosis of ovarian cancer (Log-rank=3.897 and 11.044,P=0.048 and 0.001). Conclusion:The P38MAPK signal transduction pathway was activated in ovarian cancer. The activated P38MAPK signal transduction pathway can raise the expression of uPA, which may contribute to the development of ovarian cancer. The result indicates that the P38MAPK signal transduction pathway and uPA might play an important role in the invasion and metastasis of ovarian cancer. P38MAPK and uPA might be useful markers for evaluating prognosis of ovarian cancer.

Objective To identify and investigate B(A)02 allele in a patient. Methods Serological tests were performed with standard serological methods in a patient with B(A)02 allele. DNA sequences of all seven exons and exon-intron boundaries of ABO gene were analyzed by polymerase chain reaction (PCR), direct DNA sequencing and sequencing after gene cloning. In order to analyze the allele, PyMOL software was used to establish 3D model of Glycosyltransferases B (GTB). Results The serological results showed the characteristics of B(A) phenotype. DNA analysis revealed that ABO gene of the individual was heterozygous of B(A)02/O01 allele. 700C>G mutation was identified in B101 allele, which resulted in the amino acid substitution P234A in GTB. Through the analysis of the 3D structure of GTB, it was speculated that the P234A replacement affected the intermolecular forces of the 234 amino acid and Met-266, thus changed the conformation of the donor-binding pocket of GTB,that made GTB capable of recognizing and tranferring the GalNac to the H antigen, which can lead to the formation of the weak A antigen on membrane of red blood cells. Conclusion The P234A replacement can affect the spatial conformation of the specific recognition region conformed by Met-266 and Ala-268 residues, which leads to the antigenicity change of the ABO blood group.

Objective To observe and study the influence degree of strengthen training for family members for the treatment compliance and unhealthy lifestyle of patients with diabetes mellitus. Methods A total of 64 patients with diabetes mellitus from January 2015 to March 2016 were selected and divided into two groups by the method of random number table,32 patients in control group were taken care with routine nursing of diabetes mellitus,32 patients in observation group were taken care with strengthen training for family members,then the treatment compliance,related knowledge cognition of unhealthy lifestyle and unhealthy lifestyle situation of two groups before the intervention and at 4th, 8th and 12th week after the intervention were compared. Results The treatment compliance, related knowledge cognition of unhealthy lifestyle and unhealthy lifestyle situation of two groups before the intervention all had no obvious differences (all P>0.05). The treatment compliance at 4th, 8th and 12th week after intervention were (6.26 ± 0.32), (6.95 ± 0.36), (7.28 ± 0.40) scores in observation group, and (5.23 ± 0.29), (6.01 ± 0.30), (6.30 ± 0.33) scores in control group, and the differences were significant (t=8.253, 9.201, 10.257, all P<0.05). The related knowledge cognition of unhealthy lifestyle at 4th, 8th and 12th week after intervention were (80.78±5.90), (89.44±6.02), (93.43±6.20) scores in observation group, and (73.38± 5.73), (77.93±5.84), (82.20±6.01) scores in control group, and the differences were significant (t=11.258,12.563, 13.324, all P<0.05 ). The unhealthy lifestyle situation at 4th, 8th and 12th week after intervention were 75.00%(24/32), 90.63%(29/32), 96.88%(31/32) in observation group, and 59.38%(19/32), 68.75%(22/32),78.13%(25/32) in control group, and the differences were significant (χ2=7.251, 7.468, 9.169, all P<0.05). Conclusions The influence of strengthen training for family members for the treatment compliance and unhealthy lifestyle of patients with diabetes mellitus are more active, so the value of strengthen training for family members of patients with diabetes mellitus is higher.