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.

Bile acids (BAs) are a major component of bile salt, which plays a vital role in the metabolism of lipids in humans. Ninety-five percent of bile acids are recycled by the enterohepatic circulation (EHC), and therefore EHC is essential for bile acid homeostasis. There are four transporters that mediate the transmembrane transport of bile acids, each of which plays an important role in the enterohepatic circulation. Gene defects in bile acid transporters can lead to disorders of the enterohepatic circulation, ultimately leading to clinical phenotypes such as metabolic diseases and even death. Bile transporter expression is altered in patients with various metabolic disease states, suggesting that disruption of bile acid transporters may be a pivotal pathological mechanism for the development of metabolism diseases. Thus, many drugs targeting bile acid transporters are being developed. We provide a concise overview of the progress of bile acid transporters research, discuss the relationship between different bile acid transporters and disease development, and summarize the current progress in drug development targeting bile acid transporters.

Objective To investigate the imaging characters of abdominal cocoon.Methods Six cases of abdominal cocoon proved by surgery and pathologic findings were retrospectively analyzed. Abdominal plain X-ray and CT were performed in 6 cases.The gastrointestinal barium meal series were undergone in 4 cases.The imaging findings were analyzed.Results Abdominal plain X-ray suggested intestinal obstruction in 3 of 6 cases.The gastrointestinal barium meal showed"cauliflower sign"or "concertina pattern"in all of the 4 cases;CT images revealed a conglomeration of multiple small bowel loops in all 6 cases and the intestinal loops seemed to be encapsulated in a membranelike sac.Conclusion The imaging features of gastrointestinal barium meal and CT scan could suggest the diagnose of abdominal cocoon.

Objective To explore the expression and the role of Toll-like receptor(TLR3 and TLR4) in rats with nephrotic syndrome induced by respiratory syncytial virus(RSV).Methods SD rats were inoculated intranasally and intraperitoneally with 6?106 plaque for-ming unit(PFU) RSV to construct RSV-induced nephropathy in rat model.Rats were anesthetized and blood was withdrawn from cardiac on day 4,14,30,60 after inoculation.The normal ones without intervention were set as control group.The renal histology was observed by light microscope and electron microscope.The urinary protein collected in 24 hours were measured.Meanwhile,the expressions of TLR3 and TLR4 were detected by indirect immunofluorescence staining and flow cytometry in peripheral blood mononuclear cells of rats.The results were analyzed by SPSS 13.0 softwore.Results After inoculation,the proteinuria increased and under the electron microscope the foot processes of glomerular epithelial cells were fused which resembled human minimal change nephrotic syndrome.Proteinuria reached the peak and the fusion of foot processes were most extensive in rats of RSV at 60 d.The expressions of TLR3 and TLR4 in each group of RSV-induced nephropathy in rat models were significantly higher than those in normal control group(Pa0.05).Conclusions TLR3 and TLR4 in peripheral blood mononuclear cells of RSV-induced nephropathy rat mo-dels had being significantly activated until 60 d after RSV inoculation.TLR signaling pathway may play an important role in nephrotic syndrome of rats induced by RSV.

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

AIM: To investigate the alterations of tear film after sutureless large incision manual cataract extraction (SLIMCE). METHODS: Sixty-eight SLIMCE operation eyes were studied with slit-limp microscope, break- up time (BUT), SchirmmerⅠtest (SⅠt),and fluorescence(FL) to observe the alterations of tear film at different time points in postoperation. Impression cytology and microphoto-analyses technique were also applied to observe the goblet cells at different time points postoperation(7,14,30,60,90 days). RESULTS: Subjective complaint of dry eye within 90 days after the operations were significantly increased compare with preoperations(5-27,23,19,16,13; 2-16,14,8,6,3). The schirmmer Ⅰ test were greatly increased in 14 days postoperation(10.1±4.5;15.0±4.7,13.8±5.7),the mean scores of fluorescence increased (0-17,9,5;0-8,3,1) and the mean break-up time decreased in 30 days post-operation(10.3±2.2;5.5±2.3,7.0±2.4,7.9±2.2) (P<0.05). CONCLUSION: SLIMCE operation have effect on the stability of tear film.

Ulta performance liqiuid chromatography-triple quadrupole tandem mass spectrometry ( UPLC-MS/MS) was used to monitor the relative levels of bufadienolides in toad venom in normal and bensulfuron-polluted groups. Methanol extract of toad venom was separated by UPLC ( ODS-C18 ) using a gradient elution of water contains 0. 1% formic acid and acetonitrile. Mass spectrometry was used in an ESI source operated in positive ion and MRM mode. The parameters in the source were set as follows: capillary voltage 3. 0 kV; sampling cone voltage 30 V; and desolvation temperature 500℃. In this method, external calibrations of 6 standards were typically constructed (R2=0. 9953-0. 9992). The LOD was 0. 42-4. 86 ng/mL. Intra- and inter-day precision was 3. 8%-6. 8% and 4. 0%-8. 8%, respectively. The recovery of standard was evaluated by spiking the standard compound into toad venom. Their average recoveries were 96. 9%-109. 6%, and RSDs were 2. 0%-8. 1%. This method was further employed into monitoring the levels of 36 bufadienolides. The levels of more than 20 bufadienolides were greatly different after bensulfuron pollution, suggesting that the bensulfuron pollution could change the chemical expression pattern of bufadienolides in toad venom.

Objective To establish a key technological system for spider fibroin gene code tandem connection , vector construction , prokaryotic expression and purification using genetic engineering in order to achieve MaSp 1 heterologous ex-pression in Escherichia coli and its separation and purification .Methods Isocaudarner ligation method was used to connect synthetic spider fibroin gene monomer code in tandem , and a recombinant clone concatemer was obtained .The identified recombinant clones were connected with prokaryotic expression vector pET 28a(+), and then transformed into E.coli BL21 (DE3).After being induced by IPTG for 6 hours, the expression product was identified by SDS-PAGE and Western blot-ting.Engineering bacteria were fermented in high density , and the obtained protein was purified through ammonium sulfate fractionation.Results and Conclusion The expression plasmids of MaSp1concatemers were successfully constructed , and the induced expression genetic engineering MaSp 1 protein was of the expected relative molecular mass .In addition, the pu-rity of the purified protein was above 80%.This study has developed crucial technologies for mass production of genetic en-gineering spider silk proteins .

Castleman Disease ; complications ; metabolism ; pathology ; surgery ; Dendritic Cell Sarcoma, Follicular ; complications ; metabolism ; pathology ; surgery ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Receptors, Complement 3b ; metabolism ; Thoracic Diseases ; complications ; metabolism ; pathology ; surgery ; Thoracic Wall ; Vimentin ; metabolism

Objective To evaluate the results of treating the persistent Henoch-Schonlein purpura nephritis(HSPN) by injecting methylprednisolone into intra-renal capsule in children.Methods Twenty-two patients(aged from 6 to 13 years) with persistent HSPN were randomly divided into 3 groups.Group I was treated with prednisone;group Ⅱ was treated with high dosage methylprednisolone by venous injection,while group Ⅲ was treated by injecting methylprednisolone into intra-renal capsule.The 24 h urinary protein excretion,the levels of serum albumin and creatinine,or the blood cholesterole in children with persistent HSPN were detected at the beginning,4 weeks and 8 weeks of the study.The blood pressure,body weight were detected in the study duration.Results The values of 24 h urinary protein excretion were(2.35?1.09),(0.97?0.37),and(0.99?0.52) g(P