Forty-eight patients with subclinical Cushing's syndrome(SCS)were evaluated.Eleven of them underwent adrenalectomy(Group 1)and the other 37 cases did not(Group 2).Serum and urine corticosol, plasma ACTH and parameters related to metabolic syndrome(such as waist circumference,blood pressure,blood lipids and fasting plasma glucose)were measured.The data at diagnosis were compared with those during the survey.The results indicated that patients with SCS had a significantly high prevalence of metabolic syndrome.The symptoms and signs of metabolic syndrome could be improved after removing the tumor.Otherwise there is no improvement,some patients will even develop into overt Cushing's syndrome.

Objective To explore the effects of lycium barbarum polysaccharide (LBP) on restraining the mouse pancre?atic cancer cells LTPA by the polarization of macrophages to type 1 macrophages (M1). Methods LTPA tumor model of the subcutaneous CB-17SCID mice was constructed. Model mice were randomly divided into tumor-bearing model group (n=10) and LBP treatment group (n=10). The LBP treatment group was fed 10mg/kg LBP every day, and the tumor-bearing model group was fed the same dose of normal saline. The same amount of macrophages Raw264.7 was randomly divided into the control group and experimental groups (different concentrations of LBP). MTT assay was used to detect the optical density (OD) of Raw264.7 in experimental groups and control group. ELISA was used to detect the levels of the interleukin (IL)-12 and IL-10 in experimental group (LBP was 100 mg/L) and the control group. Flow cytometry was used to test the levels of the membrane protein CD16/32 and CD206 in experimental group (LBP was 100 mg/L) and the control group. The tumor mass was weighted and the volume was calculated after three weeks. The effects of LBP on the growth of subcutaneous tumor were detected. HE staining and KI-67 staining were used to detect the microscopic changes of tumor and the proliferation of the LTPA. Results The dose of 100 mg/L LBP can promote the growth of the macrophages Raw264.7 (P<0.01), and induced the high expression of CD16/32 and low expression of CD206, high secretion of IL-12 and low secretion of IL-10. The weight, volume of the tumor and the expression of KI-67 were significantly lower in experimental group than those in the con?trol group (P<0.01). The microscopic necrosis area range of tumor was larger than that of control group. Conclusion The LBP has the effect of restraining LTPA by the polarization of macrophages to M1.

Objective To study the characteristics of Genetic typing and the antibiotic susceptibility testing of strains from Pseudomonas aeruginosa keratitis patients.Design Experimental study,Participants 23 eyes of 23 patients of Pseudomonas aeruginosa keratitis.Methods The genomic was extracted and amplified with PCR.The PCR products were purified and sequenced.The results were registered in MIST web Antibiotic susceptibility testing were performed in theses strains,Main Outcome Measures Sequence types and antibiotic susceptibility.Results The isolates were resolved into 20 STs.Two lineages were identified.MIC test showed that strains were more susceptible to aminoglycosides,The activity of quinolones and cephalosporin were higher than that of aminoglycosides.Conclusion MIST can determine homology of the strains from Pseudomonas aeruginosa keratitis by clustering results. There is no finding about relationship between Genetic typing and drug resistance.

AIM:To compare the higher-order aberrations between implantations of AcrySof and AcrySof ReStor multifocal aspherical intraocular lens(IOL) with various pupil diameters.METHODS:Fifty-four patients(62 eyes)with bilateral senile cataracts were retrospectively selected.Patients were operated with phacoemulsification and IOL implantation.They were divided into two groups based on the IOL implantation of AcrySof IQ and AcrySof ReStor.Wave front aberration:spherical aberration(C12)and the root mean square of the total higher-order aberration(RMSh)were observed 3 months after surgery.RESULTS:The larger the pupil was, the higher the C12 and RMSh were in the eyes (P<0.01).There were no statistical differences in C12 or RMSh between two groups at 5, 6 or 7mm pupil diameters (P>0.05). CONCLUSION:There are no differences between AcrySof IQ group and AcrySof ReStor group at 5, 6 or 7mm pupil diameters.

Objective:To investigate the whole genome characteristics of coxsackievirus A4 (CVA4) circulating in Qingdao city.Methods:Four CVA4 isolates circulating in Qingdao city during 2013 to 2015 were selected. Whole genome sequences of these strains were amplified by one-step reverse transcription-polymerase chain reaction (RT-PCR). Sequence alignment and phylogenetic analysis were performed using MEGA7.0 software package. Genetic recombination analysis was performed using similarity plots 3.5.1 software package.Results:Phylogenetic analysis showed that based on the sequences of the whole genome and P1, P2 and P3 regions, HS312/QD/CHN/2013 and HS605/QD/CHN/2014 strains together with the early domestic isolates belonged to the same clade, while FY218/QD/CHN/2015 strain and CV-A4/P1033/2013/China strain collected in Wenzhou in 2013 formed another clade in each phylogenetic tree. HS144/QD/CHN/2014 strain belonged to the same clade as HS312/QD/CHN/2014, HS605/QD/CHN/2014 and the early domestic CVA4 isolates in the phylogenetic tree based on the P1 region, but formed a separate clade in the phylogenetic trees based on the whole genome, P2 region and P3 region. Genetic recombination analysis revealed that there was genetic recombination between HS144/QD/CHN/2014 strain and the CVA2 strain of CV-A2/P373/2013/China isolated in mainland China in 2013 in the region of 2C-3D (5 081-7 301); FY218/QD/CHN/2015 and CV-A4/P1033/2013/China strains were highly homologous and recombination signal sequences were detected in the region of 2A-2B (3 821-4 161) between the two strains and the CVA2 strain of CV-A2/P373/2013/China.Conclusions:The CVA4 isolates circulating in Qingdao city presented obvious genetic diversity at the genome-wide level.

Objective To obtain the fusion genes of several human orphan G protein coupled receptors (oGPCRs) with Gi1α subtype of G protein and their expression system. Methods The whole open reading frames of GPR45, GPR85, GPR174 and Gilα were cloned by RT-PCR from HepG2 cDNA separately,and the corresponding fusion genes were amplified by overlap extension PCR. Then, the fusion genes-containing pBacmids were successfully constructed with the Bac-to-Bac baculovirus expression system indicated by specific transposition and virus recombination. The insect Sf9 cells were transfected with pBacmid-oGPCRs-Gi1α, and the supernatant containing recombinant virus was harvested. With the supernatant, insect Sf9 cells were infected under an optimized condition (MOI=5, infection time=72 h) and the fusion proteins were prepared and detected by Western blotting.Results The three fusion genes of GPCR45, GPR85 or GPR174 with Gi1α were obtained. The corresponding fusion proteins could be properly prepared in Sf9 cells.Conclusion Human oGPCRs could be fused with Gilα, and the fusion genes could be expressed using the Bac-to-Bac baculovirus expression system in insect Sf9 cells.

Objective To localize the sinoatrial node (SAN) of the newborn rabbits in vivo and cut it for purifying cultivation and study the morophologic characters of primary cultured pacemaker cells of SAN under light microscope and transmissional electron microscope. Methods Hearts of the newborn rabbits were embedded in paraffin for HE-staining and observed the location, form of SAN under optical microscope; SAN cells isolated from neonatal rabbits cultured and purified with the method of differential attachment and BrdU-treatment.Results SAN localized in the anterior wall of the superior vena cava and the posterior-lateral atrial wall.There was about 0.32 mm between its lowest point and sulcus terminalis. Three distinctly different types of cells were observed among the cultured cells of SAN: spindle, araneiform and polygon. The spindle cells covered the greatest proportion of the cultured cells of SAN (59.6%?7.3%). The frequency of spontaneous contraction of spindle cells was the highest among the constrcting cells (145 ?9)time/min. The results of ultrastructure observation showed that myofibrils and other organelles in spindle cells were poorly organized and significantly decreased in number compared with araneiform cells. There was no significant difference between araneiform cells isolated from SAN and from atrial muscle.Conclusion Among the cultured cells from neonatal rabbits SAN, the spindle cells are the pacemaker cells of SAN.

Objective To explore the associations between microsatellites in gonadal receptor genes and dysfunctional attitudes in adolescent with major depressive disorder(MDD).Methods Polymerase chain reaction(PCR),capillary electrophoresis and genetic scanning were performed in testing the length of microsatellites in first-onset adolescent depressive patients.Dysfunctional attitudes scale(DAS) was used in rating the dysfunctional cognition of adolescent depressive sample.These results were tested by correlative analysis and comparison analysis.Results 1.There existed significantly negative correlation between microsatellite′s length in estrogen receptor ?(ER?) gene and total score of DAS in female adolescent patients with first-onset depressive disorder.2.DAS′ total score of shorter alleles′ group was significantly higher than that of longer alleles′ group on female′ estrogen receptor ?(ER?) Gene.Conclusion The microsatellite′s length of ER? and ER? gene may have associations with dysfunctional attitudes of female adolescent with MDD.

A geldanamycin (GDM) producing strain, Streptomyces hygroscopicus 17997, was isolated from Yunnan China soil by our institute researchers. GDM is an ansamycin antibiotic, which has the ability to bind with Hsp90 (Heat Shock Protein 90) and alter its function. Hsp90 is a chaperone protein involved in the regulation of the cell cycle, cell growth, cell survival, apoptosis, and oncogenesis. So it plays a key role in regulating the physiology of cells exposed to environmental stress and in maintaining the malignant phenotype of tumor cells. As an inhibitor of Hsp90, GDM possesses potent antitumor and antivirus bioactivity, but the hypato-toxicity and poor solubility in water limits its clinical use. Two GDM derivatives, 17-(Allylamino)-17-demethoxygeldanamycin (17-AAG) and 17-dimethylamino-ethylamino-17-demethoxygeldanamycin (17-DMAG), both showing lesser hepato-toxicity, are now in Phase II and Phase I clinic trials. In order to accomplish the structure modification of GDM by genetic means, an attempt to obtain the biosynthetic gene cluster of GDM from S. hygroscopicus 17997 was made. In this study, a pair of primers was designed according to a conserved sequence of one of possible post-PKS (polyketides synthase) modification genes, the carbamoyltransferase (CT) gene (gdmN) in GDM biosynthesis. The 732 bp PCR product was obtained from the S. hygroscopicus 17997 genomic DNA. Through the colony-PCR Binary Search Method, using the CT gene primers, six positive cosmid clones, CT1-6, were identified from the S. hygroscopicus 17997 cosmid genomic library. The CT gene containing fragments were verified and localized by Southern blot. The CT-4 positive cosmid was then sub-cloned and sequenced. Approximately 28.356kb of foreign gene sequence from CT-4 cosmid and by further PCR extension reaction was obtained. Based on BLAST analysis, this sequence contains 13 possible ORFs and their deduced functions are believed to be involved in GDM production. The ORF1 encoding products show homology (87%) with incomplete sixth module and complete seventh module of PKS, gdmA3, in S. hygroscopicus NRRL 3602. The ORF2-13 gene products are similar to gdmF(9 5%), gdmM(8 8%), gdmN (92%), gdmH (92%), I (93%), J (90%), K (93%), G (96%), gdmO (91%), gdmP (93%) and two transcription regulation genes gdmRI (83%) and gdmRII (90%). The obtained possible GDM biosynthetic gene cluster in S. hygroscopicus 17997 will facilitate the further functional analysis of the genes and to modify the structure of GDM through combinatorial biosynthesis.
Anti-Bacterial Agents ; metabolism ; Benzoquinones ; metabolism ; Carboxyl and Carbamoyl Transferases ; genetics ; Chromosome Mapping ; Cloning, Molecular ; DNA Primers ; genetics ; Lactams, Macrocyclic ; metabolism ; Multigene Family ; Streptomyces ; genetics ; metabolism