Objective: To clone human apM1 gene for the study molecular mechanism of the function of adiponectin.Methods: RT-PCR method was used to isolate,from the total RNA of human adipose tissue,cDNA fragment encoding adiponectin,which was linked into pGEM-T Vector,then amplified,and cloned.The sequence was analyzed to identify the recombinant plasmids.Results: The result of sequencing indicated that the sequence of the cloned cDNA was identical to that of the corresponding parts of the mRNA for human apM1 in GeneBank except 2 nucleotides.Conclusion: Human apM1 cDNA gene was successfully cloned.

OBJECTIVE:To investigate the effect of α-interferon on long-term survival rate and tumor recurrence after radical hepatic resection. METHODS:104 patients with primary hepatocellular carcinoma underwent radical hepatic resection were divided into observation group(62 cases)and control group(42 cases). Observation group was treated with IFN-α for 18 months 4-6 weeks after surgery;control group received rountine treatment. Total survival rate and disease-free survival rate after surgery were com-pared between 2 groups. RESULTS:In observation group after surgery,1-year survival rate was 82.26%(51/62),3-year survival rate 61.29%(38/62),and 5-year survival rate 24.19%(15/62);in control group after surgery,1-year survival rate was 41.94%(26/62),3-year survival rate 28.57%(12/42)and 5-year survival rate 9.52%(4/42);there was statistically significant difference in postoperative survival rate between 2 groups (P<0.05). In observation group after surgery,1-year disease-free survival rate was 61.29%(38/62),3-year disease-free survival rate 40.32%(25/62),and 5-year disease-free surial rate 17.74%(11/62);in control group after surgery,1 year disease-free survival rate was 43.55%(27/62),3 year disease-free survival rate 22.58%(14/62),and 5-year disease-free surial rate was 0(0/62);there was statistically significant difference in postoperative disease-free survival rate be-tween 2 groups (P<0.05). CONCLUSIONS:The application of α-interferon after radical hepatic resection can significantly inhibit the recurrence of postoperative tumor,improve the long-term survival rate,and is worthy of clinical research.

Objective To study the effect of MRI on the enhancement of normal prostate, benign prostatic hyperplasia and prostate cancer by pharmacokinetic model. Methods A total of 80 patients with prostate cancer diagnosed by pathological examination in Lishui Central Hospital from February 2015 to August 2016 were selected as the subjects in this study. 1.5T superconducting magnetic resonance imaging was used, the serial needle was FMPSPGR for enhanced scan of the prostate of the patient, interlayer was 2 mm and the thickness was 7 mm, continuous scanning was performed 35 times, after the elbow vein indwelling catheter, the Gd-DTPA was injected rapidly at the beginning of the scan according to the 0.1 mmol/kg dose. By scanning data, inspection data transfer and workstation for patients with scanning results were analyzed using two room pharmacokinetic model calculation of ROI, the patients of normal prostate peripheral zone, benign prostatic hyperplasia and prostate cancer index, maximum enhancement of contrast agent distribution index, exchange rate, contrast absorption amplitude. Results After a series of calculation and analysis, the maximum enhancement index of prostate cancer, the distribution index of contrast agent, the contrast agent exchange rate and the contrast agent absorption index of prostate cancer were the highest. The maximum enhancement index, contrast agent distribution index, contrast agent exchange rate and contrast agent absorption index were significantly lower than those of prostate cancer, The difference was statistically significant (P<0.05). The indexes of normal prostate were significantly lower than those of benign prostatic hyperplasia (BPH) and prostate cancer (P<0.05). Conclusion The application of drug evaluation pharmacokinetic model of prostate MRI in different tissues to enhancing effect on patients with prostate tumor and non tumor tissue blood supply characteristics more accurately describe and differentiate, to improve the accuracy of identification and diagnosis of prostate in clinical treatment in a certain extent, with further clinical promotion and application significance.

Objective To observe the changes of microRNA141 expression in photodamaged skin of mice irradiated with ultraviolet B (UVB).Methods Eighteen C57/BL6 mice were equally divided into six groups to receive single irradiation on the hair-removed back with UVB of 0,30,60,90,180 and 270 mJ/cm2 respectively.Skin specimens were obtained from the irradiated sites 24 hours after the irradiation and subjected to paraffin embedding followed by hematoxylin-eosin (HE) staining for the observation of histopathological changes.Real-time quantitative PCR was performed to determine the miRNA141 expression in skin specimens,immunohistochemical staining (IHC) to quantify the expression of phosphatase and tensin homolog deleted on chromosome ten (PTEN) protein.The TargetScan database was employed for the prediction of miRNA141 targets,and Gostat analysis for functional clustering.Data were processed using SPSS version 13.0 software,and intergroup comparisons were done by analysis of variance.Results The relative expression level of miRNA141 in skin tissue was 2.1354 ± 0.4289,2.4333 ± 0.2517,2.9328 ± 0.3126,3.4125 ± 0.3606,4.5667 ± 0.4014 and 6.7428 ± 0.5158 in mice irradiated with UVB of 0,30,60,90,180 and 270 mJ/cm2 respectively.There was a positive correlation between the expression level of miRNA141 and radiation dose (r =0.992,P ＜ 0.01).Gostat analysis showed that the targets of miRNA141 were mainly related to transcription regulation and signaling transduction.Moreover,PTEN expression gradually decreased with the increase in radiation dose.Conclusions miRNA141 may be involved in UVB-induced photodamage and inflammatory response,and PTEN may play a regulatory role in this process.

Objective To detect the expression of translationally controlled tumor protein (TCTP) in squamous cell carcinoma (SCC) tissue and cell lines A431 and SCL-1,and to evaluate the effect of TCTP on apoptosis in and proliferation of SCC cells.Methods An immunohistochemical method was used to measure the expression of TCTP in tissue specimens from 65 patients with SCC.Western blot was performed to detect the expression of TCTP in A431 and SCL-1 cells.Three small interference RNAs (siRNAs) targeting the TPT1 gene were designed,synthesized,and transfected into A431 cells.Then,reverse transcription (RT)-PCR and Western blot were conducted to measure the expression of TPT1 mRNA and TCTP,respectively,methyl thiazolyl tetrazolium (MTT) assay and flow cytometry were carried out to detect cell proliferation and apoptosis,respectively.Results TCTP was overexpressed in SCC tissue specimens,and the expression level was positively correlated with the histologic grading of SCC (P ＜ 0.05).Western blot showed that TCTP was expressed in both A431 and SCL-1 cells,and the expression was relatively high in A431 cells.The transfection efficiency of siRNAs varied from 90％ to 95％.A decrease in the expression of TPT1 mRNA and TCTP was induced by the siRNAs in A375 cells (all P ＜ 0.05).Conclusion The downregulation of TCTP expression may increase the apoptosis in and suppress the proliferation of A431 cells.

Objective To evaluate the effects of exogenous adenovirus-mediated gene transfer of human apM1 gene (Ad-apM1) on malonaldehyde (MAD) and superoxide dismutase (SOD) levels in human umbilical vein endothelial cells (HUVEC). Methods HUVEC proliferation was measured by MTT after infection by AdapM1. Adiponetin protein level in cell culture medium of HUVEC cells infected with Ad-apM1 was detected by double antibody sandwich ELISA. The levels of MAD and SOD were measured by chromatometry. Results MTT assay showed that Ad-apMl had no effect on HUVEC proliferation. Human adiponectin protein levels in cell culture medium significantly increased after HUVEC was infected with Ad-apM1 for 24, 48, and 72 h, along with decreased MAD and increased SOD ( all P<0.05 ). MAD levels markedly increased and SOD levels decreased after HUVEC were incubated with 100 μmol/L H2O2 for 6, 12, and 24 h, and these reactions were reversed by AdapM1 transfection (all P<0.05 ). Conclusions HUVEC infected with Ad-apM1 effectively secrete human adiponectin. Ad-apM1 exerts antioxidation effect and antagonizes H2O2 -induced endothelial injury.

Aim To investigate the roles of p53-dependent signaling pathways in the process of ginsenoside Rg1 protection against 8-methoxypsoralen(8-MOP) and subsequent ultraviolet A(UVA) irradiation induced photoaging model in human dermal fibroblasts(HDFs).Methods Photoaging model was established by 8-MOP/UVA in skin HDFs.Flow cytometry, enzyme cytochemistry, immunofluorescence and Western blot were employed.Results Pretreatment with ginsenoside Rg1 could significantly reduce the amount of UVA-generated 8-oxo-dG and relieve the photoaging representation.Compared with 8-MOP/UVA treatment group, pretreatment with Ginsenoside Rg1 could decrease the expression of SA β-galatosides(SA-β-Gal), and down-regulate the level of senescence associated proteins(p16 and p21).Conclusions Ginsenoside Rg1 has prominent dose-dependent antagonism on senescence of skin HDFs induced by 8-MOP/UVA, and its mechanism may be due to its antioxidation which reduces the production of photoproducts to protect telomere against abnormal shortening.

Objective To investigate the expression changes of microRNA 146a (miRNA146a) in UVBdamaged mouse skin. Methods C57/BL6 mice were divided into dose groups to be irradiated on the back with UVB of 30, 60, 90, 180, 270 mJ/cm2 respectively for 24 hours, and time groups irradiated with UVB of 180 mJ/cm2 for 1, 12, 24 and 48 hours respectively. The mice received no irradiation served as the control. Skin samples were subsequently obtained from the irradiated sites of mice and subjected to real-time fluorescent PCR for the detection of miRNA146a expression as well as immunohistochemical staining (IHC) for the detection of STAT3. Results The expression levels of miRNA146a were 0.01158 ± 0.00098, 0.01083 ± 0.00104,0.00872 ± 0.00031, 0.00851 ± 0.00033, 0.00810 ± 0.00057 and 0.00770 ± 0.00031 in the unirradiated control mice and mice irradiated with UVB of 30, 60, 90, 180, 270 mJ/cm2 for 24 hours, respectively, 0.00730 ±0.00036, 0.00805 ± 0.00035, 0.00810 ± 0.00057 and 0.00837 ± 0.00039 in mice irradiated with UVB of 180mJ/cm2 for 1, 12, 24 and 48 hours, respectively. IHC suggested an intensive expression of phosphorylated STAT3 in mice irradiated with a high dose UVB. Conclusions miRNA146a may play an essential role in the mechanism of UVB-induced photodamge, which is mainly correlated with the negative regulation of inflammatory reaction likely mediated by the JAK/STAT3 signal transduction pathway.

Objective To explore the value of urinary albumin/creatinine ratio (ACR) in coronary heart disease(CHD) compli-cating thyroid dysfunction and its correlation with the various detection indicators of thyroid function.Methods The results of uri-nary ACR and thyroid function detection in 863 patients with CHD in our hospital from November 2015 to August 2016 were col-lected.According to the standards of American National Kidney Foundation (NKF) and Food and Drug Administration(FDA) ,863 CHD patients were divided into the group A (ACR≤10.0 mg/g · Cr) ,B(10.0 mg/g · Cr< ACR≤30.0 mg/g · Cr) ,C (30.0 mg/g · Cr300 mg/g · Cr).According to the reference values of thyroid functional indica-tors ,these patients were divided into the thyroid function normal and abnormal groups.Results The ACR level was negatively cor-related with the FT3 level in 863 CHD patients(r= -0.297 ,P<0.05) ,and was positively related with the FT 4 level(r=0.172 , P<0.05).The abnormal rate of thyroid function had significantly statistical difference among four groups (χ2 =33.533 ,P<0.05). The abnormal rate of thyroid function had significantly statistical difference between the patients with ACR >10.0 mg/g · Cr and the patients with ACR ≤ 10.0 mg/g · Cr ,the difference was statistically significant (P< 0.05) ,its risk degree was 2.083. Conclusion ACR is likely to be a risk factor for CHD patients complicated with thyroid dysfunction.It is necessary to monitor the thyroid function when ACR level >10.0 mg/g · Cr in CHD patients.

Objective To evaluate the safety of endoscopic ligation of gastric and colic full-thickness wall with nylon loops.Methods Endoscopic ligation of full gastric and colic wall with detachable nylon loops, assisted by laparotomy, were performed in 4 dogs.Ligation healing was assessed on 3, 7, 10and 17 days after the procedure.Results All animals recovered after the procedure, without any clinical signs of infection or hemorrhage.Postoperative examination did not reveal gastric or colonic perforation, or local abscess.Loop ligature showed ulcer-like lesion.Histopathology revealed muscularis propria disruption, granulation tissue bridging the site of ligation and formation of scar tissue.Regenerative mucosa coyered the surface of the ligature completely 17 days after the operation.Conclusion Endoscopic ligation of gastric and colic full-thickness wall with detachable nylon loops is safe.Its application can be extended to other fields.