To investigate the different expression of melatonin receptor between adult thyroid neoplasm and its adjacent normal thyroid tissue, we extracted the total RNA of all kinds of thyroid tissue, synthesized the primer of mt 1, MT 2, and analyzed the mRNA of melatonin receptor by RT-PCR method. mt 1, MT 2 melatonin receptor subtype positive strap expressed in thyroid adenoma as well as in adenocarcinoma. MT 2 in adenocarcinoma had a higher quantity than that in normal thyroid tissue(P0.05). Therefore, mt 1 and MT 2 subtype is present in human thyroid neoplasm tissue, and MT 2 subtype is associated with the occurrence of human adenocarcinoma.

Objective To explore the preventive effect and mechanism of pioglitazone on nonalcoholic fatty liver disease in rats.Methods Thirty six male Sprague-Dawley ( SD) rats were randomly divided into normal control group , high-fat diet group , and pioglitazone intervention group , with 12 rats in each group .Normal control group fed with standard diet for eight weeks , and the rest two groups fed with high fat diet .Pioglitazone intervention group fed with pioglitazone in the four to eight weeks , the rest two groups in the same period fed with distilled water .Fasting serum alanine aminotransferase , aspartate aminotransferase , total cholesterol , triglyc-erides, fasting glucose , and fasting blood insulin levels were determined .The fasting insulin resistance index was calculated .Routine histological features of hepatic section were observed by hematoxylin eosin (HE) staining.The shape change of Kupffer cells in the liv-er was detected .The levels of tumor necrosis factor ( TNF)-αand nitric oxide ( NO) secreted by Kupffer cells were measured .Results The fasting insulin resistance index , triglycerides , and total cholesterol in rats of high-fat diet group were significantly higher than in normal control group ( P <0.05 ) .Histopathological examination showed hepatocellular macrove sicular steatosis , lobular inflammatory cell infiltration, and inflammatory cell infiltration .Hepatic Kupffer cell morphology change , its produced TNF, NO and liver tissue pa-thology changes were positively correlated ( P <0.05 ) .The fasting insulin resistance index , triglycerides , total cholesterol was signifi-cantly lower in rats of pioglitazone intervention group than the high-fat diet group ( P <0.05 ) .The liver tissue structure was obviously improved pioglitazone intervention group rats , morphology and function of liver Kupffer cells were normal .Conclusions Pioglitazone can prevent the nonalcoholic fatty liver disease induced by high fat diet .The mechanism may be related to improve insulin resistance , reduce blood fat , and regulate function of Kupffer cells .

Objective To identify the role of autopahgy in the protective mechanism of heme oxygenase 1(HO-1) against hepatic ischemia/reperfusion (I/R) injury.Methods Forty healthy male Sprague-Dawley (SD) rats were randomly (random number table) divided into five groups (n =8 in each group),namely sham group,model group,cobalt protoporphyrin (CoPP) group,zinc protoporphyrin (ZnPP) group and 6-amino-3-methylpurine (3-MA) group.Partial hepatic I/R model was established by clamping the pedicles of left and median lobes for 1 hour and reopening for 6 hours in rats,and the rats in sham group were only received celiotomp without hepatic I/R.In the CoPP group,CoPP (a HO-1 inducer,5 mg/kg) was administered i.p 24 hours before I/R.In the ZnPP or 3-MA group,besides pretreatment with CoPP,the rats were given ZnPP (a HO-1 inhibitor,25 mg/kg) or 3-MA (an autophagy inhibitor,30 mg/kg) i.p 1 hour before I/R.Serum alanine aminotransferase (ALT) was determined with automatic biochemistry analyzer.The hepatic pathological scores (PS) were determined under light microscope using hematoxylin-eosin (HE) staining.The hepatocyte apoptosis index (AI) was assessed with terminal dexynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining.Autophagosomes in liver tissue were counted under electron microscope.The mRNA expressions of HO-1,caspase-3,Beclin-1 and Atg-5 in the liver were determined by reverse transcription-polymerase chain reaction (RT-PCR).The HO-1 activity was also measured by the generation of bilirubin with the method of double-wave spectrophotometry.Results Compared with the sham group,the level of serum ALT significantly increased in the I/R group (U/L:560.3±73.6 vs.49.1 ± 13.8,P ＜ 0.01),HE staining showed a severe hepatic injury (PS:12.0±2.0 vs.1.3±0.9,P ＜ 0.01),TUNEL staining showed a higher hepatocytes apoptosis and the expression of caspase-3 significantly increased [AI:(19.38±3.07)％ vs.(3.25±1.28)％,caspase-3 mRNA (2-△△CT):4.62±0.40 vs.1.05±0.15,both P ＜ 0.01].However,there was no significant difference in the expression of HO-1 and the genes associated with autophagy between the two groups.In the CoPP group,the hepatic injury was blunted compared with that in the I/R group [ALT (U/L):223.3 ± 34.4 vs.560.3 ± 73.6,PS:5.6 ± 2.3 vs.12.0 ± 2.0,AI:(11.38± 2.39)％ vs.(19.38 ± 3.07)％,caspase-3mRNA (2-△△CT):2.42±0.33 vs.4.62±0.40,all P ＜ 0.01].HO-1 was induced in the CoPP group and autophagy was also increased significantly after I/R when compared with those in the I/R group [HO-1 mRNA (2-△△CT):3.01 ±0.71vs.1.14 ± 0.20,HO-1 activity (pmol ·mg-1 · h-1):259 ± 37 vs.113 ± 26,the number of autophagosomes:8.75 ± 0.87 vs.1.25±0.71,Beclin-1 mRNA (2-△△CT):2.85±0.28 vs.1.15±0.11,Atg-5 mRNA (2-△△CT):2.44±0.25 vs.1.14±0.12,all P ＜ 0.01].In the ZnPP group,the activity of HO-1 was much lower than that in the CoPP group,and as a result autophagy was decreased and liver injury was increased.In the 3-MA group,although there was no difference in the activity of HO-1 compared with that in the CoPP group,autophagy was inhibited,and the protective effect of CoPP was eliminated.Conclusion HO-1 could regulate the level of autophagy during liver I/R,and in turn autophagy might mediate the protective effects of HO-1 against liver I/R injury.

Objective To optimize the technique of isolating and extracting oligosaccharide from Radix Rehmanniae.Methods The optimum conditions for the water extracting technique of Radix Rehmanniae were studied by orthogonal test design.Then the water extract was isolated by macroporous resins and active charcoal successively,meanwhile the pattern of macroporous resins,the volume of the eluents,and the decolorizing methods of active charcoal were studied.And then the oligosaccharide was obtained after being purified by Sephadex gel column chromatography.The content of stachyose was determined by HPLC.Results The optimal preparation process was as follows: 12 fold water,three times,0.5 h for each time.Then the water extract was isolated and purified by D-101 macroporous resins,0.1% active charcoal and Sephadex G15 column chromatography were successively used to obtain the oligosaccharide.The analysis of oligosaccharide indicated its main constituents are stachyose,rafinose,and manninotriose,and the content of stachyose is more than 60%.Conclusion This preparation technique of extracting oligosaccharide from Radix Rehmanniae is reasonable and feasible.And the yield of the oligosaccharide from Radix Rehmanniae achieves to 26%.

Objective To evaluate the diagnostic value of serum retinol binding protein (RBP) on the early stage of type 2 diabet‐ic nephropathy by receiver operating characteristic(ROC)curve .Methods According to urinary albumin(mAlb)/urinary creatinine (UCr) ratio ,155 patients with type 2 diabetes were divided into simple diabetic mellitus group ,early stage of diabetic nephropathy group ,and clinical stage of diabetic nephropathy group ,while healthy people were recruited randomly during the same period as con‐trol group .RBP test were performed by using immunoturbidimetry .The diagnostic value of RBP on in the early stage of type 2 dia‐betic nephropathy were evaluated by analyzing the ROC curve .Results The concentration of serum RBP in the early stage of dia‐betic nephropathy was significantly higher than that in the group of simple diabetic mellitus and control group (P<0 .05) ,the area under the ROC curve of RBP in serum was 0 .770 ,and the cutoff value was 40 .95 mmol/L ,while the sensitivity and the specificity were 81 .0% and 95 .2% .Conclusion RBP was a good marker in detecting early renal damage .

Objectives Esophageal cancer development and progression is driven by the accumulation of genetic changes. In this study, we used tissue microarray to survey the expression of p53, p16 and COX-2 in esophageal cancer progression and their clinical implication. Methods We tested p53, p16 and COX-2 proteins by immunohistochemistry and tissue microarray in 86 specimens from different stages of esophageal cancer and 40 specimens from adjacent non-cancer tissue. Results The expression of p53 and COX-2 was significantly higher in tumor tissue than that in non-tumor ones. As for the expression of pl6, no significant difference was found between tumor and adjacent tissue. An obvious relation was observed among p53, p16 and COX-2 expression that esophageal carcinogenesis was highly correlated with the positive expression of p53 or COX-2, however, no reciprocal relationship to neoplastic progression was recognized with p53, p16 and COX-2. Conclusion We observed that the tissue with the positive expression of p53 or COX-2 was more likely to develop esophageal cancer. Further work will verify the hypothesis that the expression level of p53 and COX-2 in biopsy specimen is applicable to predict early outset of esophageal cancer.

The use of optimal regulatory sequences for simultaneous expression of the transgenes might play a significant role in engineering plants with increased disease and insect resistance.The plant expression vector pOMS-GUS,which contained the GUS gene under the control of a chimeric promoter based upon the mannopine synthase(mas)promoter and the octopine synthase(ocs)enhancer,was constructed.Used as control,another vector pMAS-GUS,carried the GUS gene driven by only the mas promoter.The two vectors were introduced into tobacco plants by Agrobacterium-mediated transformation.Fluorometric assays for GUS activity and reverse transcription-polymerase chain reaction(RT-PCR)analysis revealed that GUS gene expressed weakly with untreated transgenic tobacco while the level of GUS activity increased steadily after 1 h subjected to wounding.The expression of the mas and ocs/mas promoters was induced a further 1.8-fold and 5.7-fold,respectively.SA(1 mmol/L)or MJ(250 ?mol/L)treatment also caused a large induction of the ocs/mas chimeric promoter;And the application of SA in combination with MJ(1 mmol/LSA & 250 ?mol/L MJ)produced an additive effect that exceeded the wounding response.The results showed that the ocs/mas chimeric promoter is a strong inducible promoter that can be activated by various stresses.The chimeric promoter should have utility in development of disease and insect resistant transgenic crops.

ATM: To prepare Shenfuqing Lyophilized Powder (Radix Ginsenp Rubra, Pericarpium Citri Reticulatae Viride, ect.) METHODS:Shenfuqing Lyophilized Powder was made by freeze drying. The concentration of Shenfuqing solution and the varieties and quantities of filling material were selected as well as the stability of Shenfuqing solution and powder for injection were examined primarily. RESULTS: The better prescription is 10% mannitol and 6% drug components. CONCLUSION: The quality of Shenfuqing Lyophilized Powder is more stable than that of solution for injection.

Objective:To explore the significance of detecting serum CRP,myocardial enzymes,liver function index in pediatric patients with rotavirus infectious diarrhea.Methods:110 pediatric patients with rotavirus infectious diarrhea and 85 age-matched patients with bacterial infectious diarrhea who were both treated in our hospital from January,2014 to May,2016 were enrolled in the present study.Another 20 healthy patients who had physical examination in our hospital were considered as control group.The differences of serum CRP,IL-6,hs-cTnT,CK,CL-MB,AST,ALT level and the incidence of parenteral injury were compared.Results:The incidence rate of lower respiratory tract infectivity,rash,myocardial damage and hepatic injury in RV group were significantly lower than the bacterial infectious group (P＜0.05),the level of serum CRP,IL-6 in RV group and bacterial infectious group were significantly higher than the control group,and those of RV group were significantly lower than bacterial infectious group (P ＜0.05);the level of hs-cTnT,CK,CL-MB,AST,ALT in RV group were significantly higher than bacterial infectious group and control group (P＜0.05),and those indexes of bacterial infectious group were no significantly differences compared with healthy control (P ＞ 0.05).Conclusion:The combined detection of serum CRP,myocardial enzyme and liver function indexes were of certain significance in the early distinguishing diagnosis of bacterial infectious diarrhea from rotavirus infectious diarrhea.

OBJECTIVE To investigate the contribution of the GJB6 gene [encoding connexin 30 (C?30)] mutations in Chinese population with sporadic non-syndromic hearing impairment. METHODS PCR reactions were performed with two pair of primers for the coding sequence of GJB6 gene and for the deletion of GJB6. PCR products bidirectional sequencing was subsequently applied in 214 patients with hearing loss and 86 normal controls. RESULTS A novel heterozygous mutation-233(C→A) was found, which results in amino acid change, A78D. This mutation wasn't detected in the control subjects. The altered valine residue lies within the second conserved transmembrane domain. The large deletion△(GJB6/ D13S1830)] of GJB6 was not found in this group. CONCLUSION The large deletion of GJB6 was not found in the Chinese deafness population. A novel heterozygous mutation of GJB6 was found. These results indicated GJB6 mutations are not a major cause of hearing loss in the Chinese population.