Objective To analyze the direct interaction between mmu-miR-30b and mouse FoxO3 (mFoxO3) mRNA.Methods Three target gene fragments, which were respectively 402, 123 and 299 bp in length, were amplified from mouse cDNA by PCR using specific primers and site-direct mutant primers.A complete mutant fragment was obtained by joining together the 123 and 299 bp fragments.Recombinant plasmids, pmirGLO-mFoxO3 and pmirGLO-mFoxO3 mt, were constructed through inserting wild and mutant fragments of mFoxO3 into pmirGLO vector, respectively.HEK 293T cells were respectively co-transfected with the constructed recombinant plasmids and mmu-miR-30b/mmu-miR-30b inhibitor.Dual-luciferase reporter assay system was used to determine the Firefly-Renilla luciferase activity in different groups.Western blot assay was performed to evaluate the regulatory effect of mmu-miR-30b on mFoxO3 expression.ResultsRestriction enzyme analysis and gene sequencing showed that the two recombinant plasmids, pmirGLO-mFoxO3 and pmirGLO-mFoxO3 mt, were constructed successfully.The activity of Firefly-Renilla luciferase in HEK 293T cells transfected with pmirGLO, pmirGLO+mmu-miR-30b, pmirGLO+mmu-miR-30b inhibitor, pmirGLO-mFoxO3+mmu-miR-30b, pmirGLO-mFoxO3+mmu-miR-30b+mmu-miR-30b inhibitor, or pmirGLO-mFoxO3 mt+mmu-miR-30b was 13.18±0.97, 14.35±0.99, 12.46±1.20, 9.55±1.11, 13.71±0.89 and 10.99±0.92, respectively.Compared with pmirGLO+mmu-miR-30b group, the luciferase activity was significantly decreased in pmirGLO-mFoxO3+mmu-miR-30b group (P<0.05), but showed no significant change in pmirGLO-mFoxO3 mt+mmu-miR-30b group (P>0.05).In addition, the suppressed Firefly-Renilla luciferase activity in pmirGLO-mFoxO3+mmu-miR-30b group was restored by mmu-miR-30b inhibitor treatment (P<0.05).Enhancing the expression of mmu-miR-30b could markedly inhibit the expression of mFoxO3 at protien level (P<0.05), and that could be significantly attenuated by mmu-miR-30b inhibitor treatmeat (P<0.05).Conclusion mFoxO3 mRNA is a novel target gene of mmu-miR-30b.There is a direct interaction between mmu-miR-30b and mFoxO3 mRNA.

Objective To establish a test method of dichloromethane,1,1-dichloroethylene,1,2-dichloroethylene,1,2-dichloroethane in drinking water with headspace gas chromatography.Methods Halogenated alkane hydrocarbon in the water was extracted by headspace technique,then analyzed with DB-624 capillary column.In the same time,determined with GC by controlling the temperature.Retention time of the peaks was used for qualitative analysis,while external standard method was used for quantitative analysis.Results The linear ranges for dichloromethane,1,1-dichloroethylene,1,2-dichloroethylene,trans-1,2-dichloroethylene,1,2-dichloroethane were 0.85-168,0.07-12.1,0.40-77.8,0.53-119 and 1.2-265 ?g/L respectively,the determination limits were 0.83,0.07,0.40,0.53 and 1.10?g/L respectively,r≥0.999 5,the rate of recovery were 102.5%-113.8%,and RSDs were 5.5%-11.8%.Conclusion This method is simple,rapid and sensitive,can efficiently separate and accurately determine 5 kinds of halogenated alkane hydrocarbon in the water and only takes 5.2 minutes.

Objective: To construct a high effective eukaryotic expre ss ing vector containing mcpr1 gene. Methods: mcpr1 gene w as amplified by PCR from the plasmid T-easy/ mcpr1, then PCR product was in serted into eukaryotic expressing vector pcDNA3.1/V5-His B. The positive recomb inant was identified by PCR analysis, HindIII and BamHI restriction analysis and Sequence analysis. Results: A 400 bp DNA fragment was amplified from the recombinant. Sequence analysis and restriction digest demonstrated tha t the mcpr1 gene was successfully inserted into pcDNA3.1/V5-His B plasmid. Conclusion: The eukaryotic expressed vector pcDNA3.1/V5-His B/ mcpr1 has been successfully reconstructed.

In order to enhance the students’knowledge of clinical medicine and lay the foundation for using clinical medicine,we adjusted the content and compiled pharmacognosy teaching materials,aiming at its professional features of clinical pharmacy of Xuzhou Medical College.The pharmacognosy teaching material has been applied in several rounds.Feedback from students suggested that using this teaching material was well-targeted,clear and practical.

Objective To explore the anticoagulation effect of low molecular weight heparin (LMWH) combination with warfarin at the early stage at the treatment of intracranial venous sinus thrombosis.Methods 80 cases of intracranial venous sinus thrombosis patients in people's hospital of Xinjiang Uygur autononous region from January 2010 to December 2014 were chosen to be analyzed retrospectively.37 cases in observation group were treated with low molecular heparin (LMWH) in combination with warfarin, and 43 cases in control group were treated with warfarin.The clinical curative effect between two groups was compared post-treatment.Results The effective rate in observation group was 91.89%, which was significantly higher than 67.44% in control group (P<0.05) .The recanalization rate of involved intracerebral venous in observation group was 89.19%, which was significantly higher than 67.29% in control group(P<0.05).After treatment, the prothrombin time (PT), thrombin time (TT) and activated partial thromboplastin time (APTT) were higher and fibrinogen (FBG) in both groups was lower than those pre-treatment(P<0.05).The PT and APTT was higher and FBG was lower in observation group than those in control group(P<0.05).Conclusion Low molecular weight heparin in combination with warfarin worked well than single warfarin in the treatment of intracranial venous sinus thrombosis.

Objective To observe the effects ofLinggan Wuwei Jiangxin Decoction on contents of cAMP, cAMP dependent PKA and AQP5 of model rats with clod phlegm-fluid lying latent in the lung asthma;To discuss its mechanism.Methods Ovalbumin intraperitoneal injection and atomization motivation + cold stimulation were used to establish models. Ninety rats were randomly divided into blank control group, model group, hexadecadrol group, andLinggan Wuwei Jiangxin Decoction low-, medium-, high-dose groups. All administration groups were treated with relevant medicine. The levels of cAMP, PKA, and AQP5 were detected by ELISA.Results Compared with the blank control group, the levels of cAMP, PKA, and AQP5 of rats in model group decreased obviously (P<0.05). Compared with the model group, the levels of cAMP, PKA, and AQP5 of rats in all administration groups increased (P<0.05), which increased dramatically inLinggan Wuwei Jiangxin Decoction high-dose group.ConclusionLinggan Wuwei Jiangxin Decoction can regulate the normal secretion of cAMP, PKA, and AQP5 in serum of rats with clod phlegm-fluid lying latent in the lung asthma, increase secretion of air passage liquid and decrease mucoprotein concentration, through which plays the role of treating asthma.

Objective: To establish an anti-tumor drug sensitivity test by micocapsulated tumor cells in vivo and in vitro.Methods: Alginate-polylysine-alginate(APA) microcapsule technique was used to encapsulate tongue cancer Tca8113 cells. MTT method was used to detect the cytotoxic effect of methotrexate(MTX)and fluorouracil (FU) on encapsulated cells, then IC 50 was calculated. Encapsulated cells were also seeded into the abdominal cavities of adult Kunming mice and FU was injected through tail vein. After two weeks, microcapsules were recollected and histological examination was performed . Results: Encapsulated tumor cells derived from tongue could survive and proliferate in clusters. MTX and FU inhibited the cell growth in a dose-dependant way. IC 50 of MTX and FU was 200 ?mol/L and 0.85 mg/ml respectively. Necrosis of the cells in microcapsules and fibrotic encapsulation of microcapsules were observed in the in vivo tests. There was no significant difference between treatment group and control group. Conclusion: Microcapsulted tumor cells may be used in the anti-tumor drugs sensitive experiment in vitro.

Objective To measure the expression of TGF-β1 in the sera of patients with psoriatic arthritis and its significance.Methods The serum level of TGF-β1 was measured in 38 patients with psoriatic arthritis and 80 patients with psoriasis vulgaris.Results A significant difierence was observed between the patients with psoriatic arthritis and those with psoriasis vulgaris(375.92±95.883 pg/ml vs.510.78±167.967 pg/ml,t=4.60,P<0.01).Conclusion Continuous monitoring of serum levels of TGF-β1 may help clinicians to take precautions against osteoporosis in patients with psoriatic arthritis.

Objective To observe the effect of postoperative analgesia for cesarean section with a new approach to transversus abdominis plane block.Methods 40 ASA physical status Ⅰ to Ⅱ patients undergoing cesarean section firstly were rucruited.All the patients were randomly divided into the two groups,20 cases in each group.A standard subarachnoid block was used.Group A:TAP block was performed at the end of surgery by piercing the transversus abdominis muscle from inside the abdominal wall at the midaxillary line at the level of the umbilicus with a 22-gauge blunt needle.The patients were assigned to receive 20mL of 0.298％ ropivacaine on each side of the abdominal wall.Group B:the patients were also assigned to receive normal saline.Sufentanil was used for postoperative analgesia by PICA.Each patient was assessed at 2,4,8,24 hours postoperatively using a visual analog scale,the dosage of analgesic drug and the adverse reactions.Results Compared with group B,group A had significantly lower pain scores at 2,4 hours using a visual analog scale (P ＜ 0.05) and there were no significant differernces in 8,24 hours(P ＞ 0.05).The cumulative sufentanil requirement of group A was also significantly lower at all time points (P ＜ 0.01).The incidence of sedation of the two groups were also significant differernces,eg.nausea and vomiting (15％/25％),itchy skin(5％/20％),dizziness(0/10％) (P ＜ 0.05).Conclusion This new approach to transversus abdominis plane block provides effective postoperative analgesia after cesarean section.

Objective To compare the clinical diagnoses with autopsy findings and evaluate the frequency of misdiagnosis.Methods The findings of 356 cases who were autopsied in our department due to medical treatment dispute during the period of 1988 to 2007 were retrospectively analyzed.The clinical diagnosis and autopsy findings,sex and age of the death,length of hospitalization,the hospital department,distribution of death disease and the rank of hospital were analyzed.The concordance between diagnosis before death and at autopsy was calculated.Results In 162 cases(45.5%),the autopsy findings confirmed the clinical diagnosis.In 101 cases(28.4%),the clinical diagnosis suggested by clinicians were discordant with the autopsy findings.In 63 cases(17.7%),some diagnoses made by clinicians were proved by autopsy,and in 30 cases(8.4%),the clinical and postmortem diagnosis were beyond comparison.The most frequently misdiagnosed diseases were from cardiovascular and respiratory diseases,and among them,cardiomyopathy,aortic atherosclerosis and pneumonia were most common.Conclusion Autopsy is not only helpful for the management of medical dispute,but also beneficial to reduce the misdiagnosis in clinical practice.