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.

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 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.

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.

Human bocavirus (HBoV) 1-4 have been detected both in respiratory and stool samples since the first HBoV was discovered in 2005. HBoV-1 is mostly associated with respiratory infection, while HBoV 2-4 are usually associated with intestinal tract infection. A variety of signs and symptoms have been described in patients with HBoV infection, including cough, wheezing, pneumonia, and diarrhea, but the research on pathogenic mechanism of HBoV is limited because HBoV cannot be cultured in vitro due to the lack of appropriate host cells. Three-dimensional epithelial cell culture, reverse genetics, and viral metagenomics are identified as novel tools that may promote the research on pathogenic mechanism of HBoV and the discovery of new viruses. This review summaries currently available diagnostic approaches such as electron microscopy, cell culture, PCR, and immunoassay in order to provide a method reference for indepth research on HBoV.
Animals ; Human bocavirus ; genetics ; growth & development ; isolation & purification ; pathogenicity ; Humans ; Parvoviridae Infections ; diagnosis ; virology ; Viral Proteins ; genetics ; metabolism ; Virology ; methods ; Virulence ; Virus Cultivation

Objective To investigate the clinical and endoscopical features of patients suffered from Henoch-Schonlein purpura (HSP). Methods Retrospective analysis of the clinical manifestation in 197 purpura patients, of which 81 cases were HSP, and described the endoscopic features of 19 cases who underwent gastroscopy and/or colonoscopy. The specimens from 15 cases were evaluated histopathologically. Results Eighty-one patients were diagnosed as HSP, 51 (63. 0% ) of them had a history of upper respiratory tract infection 1 -3 weeks before, taking antibiotics or animal protein prior to the onset of the illness. In 21 patients, abdominal symptoms presented firstly, which occurred 1 -40 days prior to the appearance of skin rashes. Seventy- four (91. 4% ) patients experienced abdominal pain, 37(45. 7% ) patients had digestive tract bleeding. Endoscopic features included diffused congestive edema of gastrointestinal mucosa, widespread hemorrhagic spots, erythema, erosion and ulceration. Lesions were relatively severer in small intestine than those in large intestine. Histological manifestations showed massive neutrophilic infiltration in mucosa and submucosa, fibrotic necrosis of small vessels, focal bleeding, erosion and ulceration. There was prominent accordance in the extents of endoscopic and pathologic manifestation with the severity of gastrointestinal symptoms. Conclusions Forty one percent of purpura patients presented as HSP, of them 26% had the abdominal symptoms firstly. Small bowel lesions were severer than those of stomach or colon. The typical features of the illness and endoscopic findings are very helpful to the early diagnosis of HSP.

Objective To investigate the change and clinical significance of serum hepcidin,serum ferritin (SF),transferrin re-ceptor (sTfR)and serum iron (SI)in patients in type 2 diabetes(T2DM).Methods 130 patients with T2DM were divided into 2 groups according to the 24 hour urine microalbumin (mAlb)quantitative:group A for trace microalbumin group 45 ca-ses (mAlb30~300 mg/24 h),group B for normal albuminuria group of 85 cases,an alternate period of 45 cases of healthy physical examination for group C (control group).Results Serum hepcidin and SF of group A (42.27±32.12 ng/ml,211.6 ±107.2 ng/ml)were significantly higher than those in group B (26.12 ± 18.36 ng/ml,179.1 ± 109.7 ng/ml)and the healthy control group (P <0.05),hepcidin and SF of group B was significantly higher than that of the control group (9.47 ±1.65 ng/ml,84.41±47.10 ng/ml,P <0.01),SI and transferrin receptor(sTfR)has no statistical significance between the three groups (P >0.05).Correlation analysis showed that patients with type 2 diabetes hepcidin was positively related with SF (P <0.05),hepcidin and sTfR,SI had no significant correlation.Conclusion These results indicated that there existed serum hepcidin and SF increased iron overload and iron metabolism disorder in type 2 diabetes.Therefore,detection of serum iron and SF can be used as a predictor of diabetes early renal damage.