Objective To construct and appraise a series of shRNA eukaryotic expression plasmid which contains vary spots of human SKP2 gene.Methods Three pairs of hairpin-like oligonucleotide sequences specific for human SKP2 gene were designed and synthesized. They were cloned into the eukaryotic expression pSIREN-RetroQ plasmid. The recombinated plasmids were confirmed by PCR and sequencing.Results The shRNA sequences were constructed successfully, and could be inserted into the eukaryotic expression vector pSIREN-RetroQ.Conclusions Three shRNA transcripting plasmids pSIREN-RetroQ are successfully constructed, and form a basis for research in cancer gene target therapy against SKP2 gene.

Polyketides are very large group of natural products with functional and structural diversity.Most of them are produced by microor- ganism and have medicinal activities,including antibiotic,anticancer,antifungal and antiparasitic properties.The researchs in this area have progressed greatly.More and more polyketides are discovered,on the other hand the mechanisms of biosynthesis of those various polyketides are researched more deeply and clearly.The article reviewed the progress of the research in the diversity of polyketide synthases and the mechanisms of polyketide biosynthesise.

OBJECTIVE:To observe the effects of parecoxib preemptive analgesia combined with postoperative epidural analgesia on hip replacement and immune function. METHODS:Totally 86 patients undergoing unilateral hip replacement selected from Xining Third People's Hospital during Jan. 2014-Jun. 2016 were divided into observation group and control group according to random num-ber table,with 43 cases in each group. Two groups underwent unilateral hip replacement under spinal epidural anesthesia. Observation group was given Parecoxib sodium for injection 40 mg intravenously 30 min before the end of surgery. Control group was given con-stant volume of normal saline intravenously. Both groups were given epidural analgesia pump(Ropivacaine hydrochloride injection 150 mg+Ondansetron hydrochloride injection 16 mg+Dezocine injection 10 mg,added into normal saline to 100 mL)after surgery with dripping speed of 2 mL/h and patient-controlled time of 15 min. The hemodynamic indexes,analgesic effect,T lymphocyte subsets and NK cells levels were compared between 2 groups at different time points as well as the occurrence of ADR after surgery. RE-SULTS:Two patients of observation group and one patient of control group withdrew from the study. Before surgery,there was no sta-tistical significance in MAP and HR between 2 groups(P>0.05);12,24 h after surgery,MAP and HR of control group were signifi-cantly increased and higher than observation group,with statistical significance(P<0.05). There was no statistical significance in MAP and HR of observation group at different time points compared to before treatment;there was no statistical significance in MAP and HR between 2 groups immediately after surgery,48 h after surgery(P>0.05). There was no statistical significance in VAS score between 2 groups immediately after surgery(P>0.05);6,12,24,48 h after surgery,VAS scores of observation group were signifi-cantly lower than those of control group,with statistical significance(P<0.05). There was no statistical significance in CD3+,CD4+, CD4+/CD8+and NK cells levels between 2 groups before surgery(P>0.05). CD3+,CD4+and NK cells levels of 2 groups immediately after surgery,CD3+,CD4+,CD4+/CD8+ and NK cells levels of 2 groups at 12,24 h after surgery,CD3+and CD4+of control group at 48 h after surgery all changed significantly;CD3+,CD4+,CD4+/CD8+and NK cells levels of observation group at 12,24 h after sur-gery were significantly higher than those of control group,with statistical significantly(P<0.05). There was no statistical significance in the incidence of ADR between 2 groups(P>0.05). CONCLUSIONS:The parecoxib preemptive analgesia combined with postopera-tive epidural analgesia shows good analgesic effect for hip replacement patients,keeps their hemodynamics stable and protects immune function to certain extent with good safety.

Objective:To explore side population(SP) cells in human epithelial cells and to observe the expression of the universal stem cell marker ABCG2,epidermal stem cell markers ?6 integrin and ?1 integrin on SP cells.Methods:Epithelial cells were obtained by digesting human skins with Dispase II and Trypsin and stained with Hoechst33342 and PI.The SP cells were analyzed and sorted by the fluorescence-activated cell sorter.Then the expression of ABCG2,?6 integrin and ?1 integrin was analyzed by flow cytometry.Results:SP cells accounted for 0.2%-0.3% of total human epithelial cells.Only a small part of cells expressed ABCG2,integrin ?6 and integrin ?1 of both SP cells and total human epithelial cells detected by flow cytometry.The difference of positive rates between SP cells and total human epithelial cells was not significant.Conclusion:SP cells accounted for 0.2%-0.3% of total human epithelial cells.The difference of positive rates of stem cells marker ABCG2,integrin ?6 and integrin ?1 between SP cells and total human epithelial cells was not significant.

Objective To analyze the clinical,radiological and pathological features of primary leiomyosarcoma of bone and to im-prove the diagnostic level.Methods Totally 6 cases of PLB proved by pathology were collected in our study.The clinical,radiologi-cal and pathological findings were retrospectively analyzed,and combined with the relevant literature to have a comprehensive analy-sis.Results The lesions of 6 patients were solitary,located in femoral (1 case),distal femur (1 case),proximal tibia (2 cases),is-chium (1 case)and humerus (1 case).There was no periosteal reaction in all 6 cases but they all had soft tissue mass,sometimes there may have some mineralization within the tumor.Ill-defined osteolytic bone destruction was detected in 5 patients,while expan-sion of cystic bone destruction was found in the other 1 patients.3 cases had hardened edge,2 cases with adjacent bone cortical thickening.The MR imaging in 5 patients showed iso-intensity signal on T1 WI,a slightly inhomogeneously higher signal on T2 WI and a significant heterogeneously enhancement after injected of Gd-DTPA.Another 1 patient showed a double ring sign,the central area onT1 WI and T2 WI expressed heterogeneous slightly higher signal,the peripheral annular zone was iso-intensity on T1 WI and slightly higher on T2 WI,after injected contrast medium,it was inhomogeneous enhancement.In immunohistochemically,all tumors reacted positively with SMA,3 patients with Desmin and Vimentin positive,2 patients with Ki-67 positive,and there had some scat-tered cells with CD68 positive,but the S-100 were all negative.Conclusion The clinical,radiological and conventional histological manifestations of PLB are not specific.Immunohistochemical and/or ultrastructural analysis who displayed smooth muscle differenti-ation can confirm the diagnosis.

ObjectiveTo investigate whether aldosterone infusion induces glomerular or podocyte injury in rats and to evaluate the effects of eplerenoen (EPL), andodipine (CCB) and telmisartan (ARB) on aldosterone- induced injury. MethodsThirty male Sprague-Dawley rats were divided into 5 groups: control, subcutaneous infusion of aldosterone (1.5 μg/h, ALD group) and aldosterone infusion plus eplerenone (100 mg·kg-1·d-1, EPL group), amlodipine(10 nag·kg-1·d-1 CCB group), telmisartan (3 mg·kg-1·d-1, ARB group), respectively. Systolic blood pressure(SBP) and urinary albumin excretion ratio(UAER) were measured at day 0, 7, 14, 21, 28. Blood samples were harvested to detect plasma angiotensin Ⅱ, plasma aldosterone, serum sodium, serum potassium and serum creatinine at day 28. Glomerular damge was quantified by morphological glomerular injury score (GIS). Immunohistochemistry and RT-PCR were performed to evaluate podocyte lesion, and apoptosis ratio of pedocyte (ARP) in a glomerular cross section was analyzed by TUNEL. ResultsALD infusion progressively increased SBP and UAER compared with CTL (P＜0.01). SBP was significantly reduced in EPL, CCB or ARB-treated animals, meanwhile, UAER was decreased in EPL and ARB group, but not in CCB group. The ALD-infused rats exibited hypernatremia and hypopotassaemia, which were blocked by EPL adminstration but not by CCB or ARB treatment. ARB group had a significant increase in plasma angiotensin Ⅱ compared with ALD, CCB and EPL groups(P＜0.01). The ALD-infused animals developed hyperaldosteronemia compared with CTL, but with no difference of plasma aldosterone among ALD, EPL, CCB and ARB-treated rats. Treatment with EPL prevented an increase of GIS and ARP compared with CCB and ARB (P＜0.05, P＜0.01). Protein and mRNA expression of nephfin was up-regulated in ALD group (P＜ 0.01), but was significantly prevented by EPL treatment(P＜0.01), whereas CCB and ARB therapy had no such effect. Conclusion ALD infusion significantly induces glomerular and pedocyte injury which is blocked by EPL but not by CCB or ARB independently on systemic hemodynamics.

Objective To evaluate the accuracy of preoperative MRI in the diagnosis of malformations associated with MRKH syndrome and identification of uterine endometrium to optimize the clinical management.Methods 1 8 females with primary amenor-rhea were studied with MRI performed with a 1.5T or 3.0T imager.All patients were examined in the supine position using a phased-array coil and underwent pelvic MRI.Two experienced radiologists evaluated all the examinations in consensus to assess the pres-ence,position and morphology of vagina,uterus,ovaries and any pelvic abnormalities.Results One or two rudimentary uteri were identified in 17 patients (94.4%).A total of 16 patients (88.9%)had bilateral rudimentary uteri,1 (5.6%)had unilateral rudimentary uteri(it was left sided).1 (5.6%)had no uterine remnants.4 (12.1%)of 33 uterine buds in 4 patients showed differentiation of the center of the uterus into three layer.Bilateral ovaries were present in 1 7 patients,and their shapes,sizes and signals are normal.The vagina and cervix were absent in all cases.Conclusion MRI is a useful diagnostic tool in the preoperative evaluation of MRKH syndrome and the endometrium to further optimize the treatment plan.

Objective To observe whether dermal multipotent stem cells (dMSCs) treated with platelet-derived growth factor-AA ( PDGF-AA )could distribute more frequently to the bone marrow in rats of total body irradiation (TBI).Methods Male dMSCs were isolated and 10 μg/L PDGF-AA was added to the culture medium and further cultured for 2 h.Then the expression of tenascin-C were examined by Western blot, and the migration ability of dMSCs was assessed in transwell chamber.The pre-treated dMSCs were transplanted by tail vein injection into female rats administered with total body irradiation, and 2 weeks after transplantation, real-time PCR was employed to measure the amount of dMSCs in bone marrow.Non-treated dMSCs served as control.Results PDGF-AA treatment increased the expression of tenascin-C in dMSCs, made (1.79 ± 0.13) × 105 cells migrate to the lower chamber under the effect of bone marrow extract, and distributed to bone marrow in TBI rats, significantly more than ( 1.24 ± 0.09) ×105 in non-treated dMSCs (t = 8.833, P ＜ 0.0l ).Conclusions PDGF-AA treatment could enhance the migration ability of dMSCs and increase the amount of dMSCs in bone marrow of TBI rats after transplantation.

Objective To investigate the effect of norcantharidin on growth inhibition and induction of apoptosis of human rectal cancer Colo 320 cells. Methods Norcantharidin (NCTD) in different concentrations were added to rectal cancer Colo 320 cells. Morphological characteristics of apoptosis were observed using the light microscope and transmission electron microscope. The expressions of Bag-1 and Bcl-2 proteins were tested by Western blotting. The growth inhibition of Colo 320 cells on the cell cycle was observed by flow cytometry. Results The apoptosis morphological changes of Colo 320 cells were observed by the light microscope and transmission electron microscopy. Flow cytometry analysis showed that the cell count of G2/M phase in experimental group was higher than that in control group ( <0.05) but the cell counts of G0/G1 and S phases have decreased in experimental group after treatment with NCTD at the concentrations of 5μg/mL, 10μg/mL and 20 μg/mL, and presented dosage dependence relations. The expressions of Bag-1 and Bcl-2 proteins have decreased. Conclusion Norcantharidin has inhibitory effect on rectal cancer Colo 320 cells, and the effect may be related to the cell cycle arrest and apoptosis.

Phosphorylation is the most common way of p 53 post-translational modifications .However , gaps still exist in our knowledge regarding the role and mechanism of phosphorylation of p 53 at Ser392 in carcinogenesis and cancer prevention.In the present study, we summarized the effect of phos-p53-Ser392 to wild-type and mutant p53, the regulation by DNA damage agents and protein kinase , and the significance of phosphorylation of p 53-Ser392 in cancer treatment .