Objective:To establish a random amplified polymorphism DNA(RAPD) method for classification of origin fungi of the Chinese traditional medicine Red Kojic. Methods:The genome DNA of 7 ordinary species Monascus including M. purpureus and a strain of Aspergillus terreus were extracted by CTAB. The content of DNA was assayed doubly by fluorescence intensity of ethidium bromide (EB) and spectrometer. Amplification products were detected by agarose gel electrophoresis and clustering analysis by PHYLIP 3.5c. Results:A characteristic pattern was produced depend on one of the 10 primers screened from 60 random primers, and the genome was amplified. The diversities of the fingerprint patterns of Monascus was obvious, by which Monascus can be distinguished easily. The results were consistent with that of morphologic study. Conclusion:RAPD is a new assay technique. It can be used to classify and identify of Monascus fungi. [

Objective To observe the effect of residual tumor after radiotherapy on survival rate of patients with nasopharyngeal carcinoma (NPC). Methods From Jan. 1989 to Dec. 1998, 108 of 304 NPC patients pathologically confirmed, had residual tumor after radical radiotherapy, of whom, 26, 68, 14 had residual lesion in the nasopharyngeal cavity alone, cervical lymph node alone, and nasopharyngeal cavity plus cervical lymph node. Results The overall 1 , 3 , 5 , and 10 year survival rates (OS) significantly decreased in the residual group. The highest OS was in the group with nasopharynx residual alone, and the lowest in nasopharynx plus residual lymph node group. The bigger the residual lesion, the lower the OS. Conclusions The overall survival rate decreases in patients with residual lesion after radiotherapy, especially in patients with both nasophrynx and regional lymph node residual. Tumor residual after radiotherapy can be a prognostic indicator for patients with NPC.

Objective：To establish a capillary gas chromatography(CGC) method for identifying the origin fungi of traditional Chinese medicine Hongqu. Methods：The volatile components of 7 species of Monascus including M. purpureus Went, M.aurantiacus Lee, M. serorubesceus　Sato, M. albidus Sato, M. barkeri Dangerd, M. ruber van　Tieghem, and　M.fuliginosus　Sato were analyzed by capillary gas chromatography method. Inlet temperature and detector temperature were 280℃，temperature program were 130℃,5 min→10℃/min→200℃, 10 min FID was used for detection. Results：The variety and contents of the volatility components in Monascus had obvious differences which can be distinguished easily by the main fingerprint peaks within 20 min. Conclusion：This method is useful to identify of Monascus fungi.

BACKGROUND:Establishment of in vitro culture system of human placental microvascular endothelial cel s with high purity is very important. In recent studies, some scholars have successful y obtained a large number of placental microvascular endothelial cel s by three-stepenzyme digestion and magnetic separation method, but the procedures were extremely complex and it had great damage to the cel s. Therefore, how to separate human placental microvascular endothelial cel s easily and obtain high-purified cel s has become a research hotspot.
OBJECTIVE:To investigate an efficient method to isolate and purify human placental microvascular endothelial cel s from early vil us microvessels, observe the cel growth and identify the cel s.
METHODS:The vil i from normal early pregnancies (6-8 weeks) after artificial abortion were col ected aseptical y. Using two-step digestion procedure and discontinuous Percol density gradient centrifugation method, human placental microvascular endothelial cel s were obtained. Then the cel s were identified by trypsin digestion method and repeated adherence method.
RESULTS AND CONCLUSION:Human placental microvascular endothelial cel s were isolated successful y from early vil i. The primary cel s adhered to the wal s after inoculated for 24 hours and entered logarithmic phase at 10 days. 80%of the cel s achieved a confluence at 12-13 days after inoculating. The subculture cel s grew swiftly with the typical cobblestone appearance. Immunofluorescence staining showed that, cultured human placental microvascular endothelial cel s demonstrated a strong positive reaction to von Wil ebrand factor antigen and CD31, accounting for 100%. MTT assay results showed that, human placental microvascular endothelial cel s at passage 5 exhibited an S-shaped growth curve. High-purity human placental microvascular endothelial cel s can be obtained by proteolytic enzymes digestion and discontinuous Percol density gradient centrifugation method, and the purity is detected by trypsin digestion method and repeated adherence method.

Objective To explore the influences of the comprehensive nurse-doctor collaboration intervention on sleep quality of the lung cancer patients receiving chemotherapy. Methods A total of 94 lung cancer patients receiving chemotherapy were selected from October 2012 to December 2013 and equally randomized into the observation group and control group, with 47 patients in each group. The patients of control group were treated with routine nursing for lung cancer. The patients of observation group adopted the “nurse-doctor collaboration” management model including psychotherapy, relaxation therapy, sleep restriction therapy,
adjust treatment on the basis of routine nursing. After four weeks intervention, the effects were evaluated according to the sleep quality of patients. Results A total of 30 patients could sleep well in the observation group whereas its number was 17 in the control group. The observation group and control group effectiveness were 63. 8% and 37. 8% respectively. The effect of sleep quality in observation group was significantly better than the control group (χ2 =7. 191,P=0. 007). Conclusions The comprehensive nurse-doctor collaboration measures can obviously correct the bad sleep habits and sleep behavior in lung cancer patients with chemotherapy, and improve the quality of sleep in those patients.

Objective To evaluate the diagnostic efficacy of Kwak and ACR( 2017 ) thyroid imaging reporting and data systems ( T I‐RADS ) for thyroid nodules . Methods Cases of thyroid nodule who underwent surgery from January 2015 to M arch 2018 in 15 hospitals in Sichuan province were collected and the ultrasonographic features of thyroid nodules were retrospectively analyzed by trained senior ultrasound physicians using Kwak and ACR T I‐RADS classification methods . Totally ,12 712 thyroid nodules were observed ,7 023 thyroid nodules in 7 023 cases with complete ultrasound and surgical and pathological data were eventually enrolled in the study . T hyroid nodules with solid ,hypoechoic or very hypoechoic ,tall/wide ratio ≥ 1 , margin ill‐defined and microcalcification were classified as malignant signs of ultrasound . M alignant percentage was calculated and diagnostic tests were performed . Results ① T here was a statistical difference between the benign and malignant nodules in the two types of T I‐RADS classification ( P＜0 .01) . ② T he area under ROC curve of Kwak and ACR in the diagnosis of malignant nodules were 0 .89 and 0 .84 ,respectively . T he Youden index of Kwak and ACR were 0 .66 and 0 .57 ,respectively . ③Taking Kwak T I4B and ACR T R4 as critical points for malignancy ,the sensitivity ,specificity ,positive predictive value and negative predictive value of Kwak T I 4B were 75 .0% ,90 .9% ,83 .2% ,and 85 .9% , respectively . T he accuracy of Kwak T I4B was 84 .9% ; T he sensitivity ,specificity ,positive predictive value and negative predictive value of ACR T R4 were 88 .2% ,68 .9% ,62 .9% ,and 90 .8% ,respectively . T he accuracy of ACR T R4 was 76 .2% . T he Kappa value of Kwak TI4B and ACR T R4 was 0 .52 . T he χ2 value of Kwak T I4B and ACR T R4 was 2 174 .6 ( P ＜ 0 .01 ) . Conclusions T he diagnostic values of two T I‐RADS classification methods for thyroid malignant nodules are high . T he overall efficiency of Kwak T I‐RADS classification method is better than that of ACR TI‐RADS classification method .

Benign prostatic hyperplasia(BPH)is one of the major chronic complications of type 2 diabetes mellitus(T2DM),and sex steroid hormones are common risk factors for the occurrence of T2DM and BPH.The profiles of sex steroid hormones are simultaneously quantified by LC-MS/MS in the clinical serum of patients,including simple BPH patients,newly diagnosed T2DM patients,T2DM complicated with BPH patients and matched healthy individuals.The G protein-coupled estrogen receptor(GPER)inhibitor G15,GPER knockdown lentivirus,the YAP1 inhibitor verteporfin,YAP1 knockdown/overexpression lentivirus,targeted metabolomics analysis,and Co-IP assays are used to investigate the molecular mechanisms of the disrupted sex steroid hormones homeostasis in the pathological process of T2DM complicated with BPH.The homeostasis of sex steroid hormone is disrupted in the serum of patients,accompanying with the proliferated prostatic epithelial cells(PECs).The sex steroid hormone metabolic profiles of T2DM patients complicated with BPH have the greatest degrees of separation from those of healthy individuals.Elevated 17β-estradiol(E2)is the key contributor to the disrupted sex steroid hormone homeostasis,and is significantly positively related to the clinical characteristics of T2DM patients complicated with BPH.Activating GPER by E2 via Hippo-YAP1 signaling exacerbates high glucose(HG)-induced PECs prolifer-ation through the formation of the YAP1-TEAD4 heterodimer.Knockdown or inhibition of GPER-mediated Hippo-YAP1 signaling suppresses PECs proliferation in HG and E2 co-treated BPH-1 cells.The anti-proliferative effects of verteporfin,an inhibitor of YAP1,are blocked by YAP1 overexpression in HG and E2 co-treated BPH-1 cells.Inactivating E2/GPER/Hippo/YAP1 signaling may be effective at delaying the progression of T2DM complicated with BPH by inhibiting PECs proliferation.